KR20170078561A - Novel genes for regulating the virulence of Cryptococcus neoformans and their use - Google Patents
Novel genes for regulating the virulence of Cryptococcus neoformans and their use Download PDFInfo
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Abstract
본 발명은 크립토코커스 네오포만스 균주의 독성을 조절하는 신규 유전자 및 이의 용도에 관한 것이다. 본 발명에 따라 크립토코커스 네오포만스 균주의 신규 전사 인자의 발현 수준을 탐지하여 항균 효과 또는 뇌수막염 치료 효과를 가지는 후보물질을 효과적으로 탐색할 수 있다. 또한, 기존의 항균제 또는 뇌수막염 치료제와 병용 투여시 상승효과를 가질 수 있는 후보물질에 대한 스크리닝 방법을 제공하며, 신규한 전사 인자의 발현을 증가 또는 억제하여 항균 효과 또는 뇌수막염 치료효과가 있는 약학 조성물을 제공할 수 있다.
본 발명에서 곤충 모델 및 동물 모델을 이용한 대규모의 독성 분석을 실시하여 크립토코커스 네오포만스의 병원성에 관련된 다수의 전사인자를 확인하였고, 균질 병원균의 생체 외 및 생체 내 표현형 특성 사이의 복잡한 상관관계를 규명하였다. The present invention relates to a novel gene which regulates the toxicity of Cryptococcus neoformans strains and uses thereof. According to the present invention, it is possible to effectively detect a candidate substance having an antibacterial effect or an effect of treating meningitis by detecting the expression level of a novel transcription factor of Cryptococcus neoformans strain. The present invention also provides a screening method for a candidate substance which can have a synergistic effect when used in combination with a conventional antimicrobial agent or a therapeutic agent for meningitis, and provides a pharmaceutical composition having an antimicrobial effect or a therapeutic effect for meningitis by increasing or inhibiting the expression of a novel transcription factor .
In the present invention, a large-scale toxicity analysis using insect models and animal models was conducted to identify a number of transcription factors related to the virulence of Cryptococcus neoformans, and a complex correlation between the in vitro and in vivo phenotypic characteristics of homologous pathogens Respectively.
Description
본 발명은 크립토코커스 네오포만스 균주의 독성을 조절하는 신규 유전자 및 이의 용도에 관한 것이다. 또한, 본 발명은 크립토코커스 네오포만스 균주의 독성 조절에 관련된 유전자의 발현을 확인하는 것을 특징으로 하는 항진균제 스크리닝 방법 및 뇌수막염 치료용 약제의 스크리닝 방법에 관한 것이다.The present invention relates to a novel gene that modulates the toxicity of Cryptococcus neoformans strains and uses thereof. In addition, the present invention relates to an antifungal agent screening method and a screening method for a drug for the treatment of meningitis, characterized in that the expression of a gene related to the regulation of toxicity of a Cryptococcus neoformans strain is confirmed.
진균 감염은 건포상백선, 완선, 아구창 같은 국소적 감염이 많이 발생하였고, 전신적 진균 감염은 드물게 발생했지만, 최근에는 전체 병원 내 감염에서 높은 빈도를 차지할 정도로 흔하게 발병하고 있다. 현재까지 개발된 항진균제는 화학적 구조에 따라 크게 아졸(azole) 구조를 갖는 항진균제와 아졸 구조를 갖지 않는 항진균제로 분류할 수 있다. 아졸 계열의 항진균제로 케토코나졸(ketoconazole), 플루코나졸(fluconazole), 이트라코나졸 (itraconazole), 보리코나졸(voriconazole) 등이 있고, 비-아졸 계열의 항진균제로 테르비나핀(terbinafine), 플루사이토신(flucytosine), 암포테리신 B(Amphotericin B), 카스포푼긴(caspofungin) 등이 있다. 아졸 구조를 갖는 케토코나졸, 플루코나졸, 이트라코나졸, 보리코나졸과 알릴아민(allylamines) 계열인 나프티핀, 테르비나핀은 유사한 작용기전을 지니고 있다. 두 계열의 항진균제는 라노스테롤이 진균 세포막의 주성분인 에르고스테롤로 전환되는 과정에 필요한 효소를 억제하는 작용을 나타낸다. 아졸 계열 항진균제는 미세소체 효소를 억제하고, 알릴아민 계열의 항진균제는 스쿠알렌 에폭시데이즈(epoxidase)를 억제하여 위와 같은 효과를 나타낸다. 플루사이토신(5-FC)은 핵산합성을 억제하는 대사길항제로서 진균 RNA의 오부호전달 유발 및 DNA 합성을 비경쟁적으로 길항하여 항진균작용을 나타내며, 폴리엔(Polyenes) 구조를 가진 암포테리신 B는 진균 세포막 내부의 에르고스테롤에 결합하여 세포막의 탈분극을 유발하고, 구멍을 형성하여 세포 내 함유물의 손실을 유발하여 항진균 작용을 나타낸다. 에키노칸딘(Echinocandins) 계열의 항진균제인 카스포푼긴은 진균 세포벽 형성을 가역적으로 억제하는 작용을 지니고 있으며, 세포벽에 작용한다는 점에서 위에 언급한 세포막에 작용하는 항진균제와 차이가 있다. 아졸 계열의 약물은 간기능 저하 환자에게 사용시 간염에 의한 사망을 초래할 수도 있으므로 투여 전에 반드시 간기능 검사가 선행되어야 한다. 플루사이토신은 용량의존적으로 골수 억제 작용, 간독성이 나타나고 소장결장 염이 발생 가능한 것으로 보고되었고, 이런 부작용은 신기능이 저하된 경우 더 증가하므로 환자의 신기능 모니터링이 매우 중요하다. 또한 임산부에서는 사용이 금지되어 있다. 암포테리신 B의 대표적 독성은 신동맥 수축에 따른 사구체 신독성으로, 용량 의존적이어서 평생 누적 용량이 4~5g인 경우 영구적인 신기능 손실 발생률이 증가한다. 또한, 세뇨관 독성에 의한 칼륨, 마그네슘, 중탄산염의 과도한 소실 및 조혈호르몬 생산 저하 등의 신독성이 일어날 수 있다. 그 외 급성 반응으로 혈전정맥염, 오한, 떨림, 과호흡 등의 증상이 나타날 수 있다.Local infections such as ringworm ringworm, jock itch, and thrush were common, and systemic fungal infections were rare, but recently, they are common enough to account for a high frequency in all hospital infections. Antifungal agents developed so far can be classified as antifungal agents having an azole structure and antifungal agents not having an azole structure according to their chemical structure. The azole-based antifungal agents include ketoconazole, fluconazole, itraconazole, and voriconazole, and the non-azole-based antifungal agents terbinafine and flucytosine , Amphotericin B, and caspofungin. Ketoconazole, fluconazole, itraconazole, voriconazole and allylamines having an azole structure, naphthipine and terbinafine, have similar mechanisms of action. Both classes of antifungal agents show the action of inhibiting enzymes required for the process of converting lanosterol to ergosterol, the main component of the fungal cell membrane. Azole-based antifungal agents inhibit microsomal enzymes, and allylamine-based antifungal agents inhibit squalene epoxidase, thus exhibiting the above effects. Flucytosine (5-FC) is a metabolic antagonist that inhibits nucleic acid synthesis, and exhibits antifungal action by inducing miscoding of fungal RNA and noncompetitively antagonizing DNA synthesis. Amphotericin B with polyenes structure Is bound to ergosterol inside the fungal cell membrane to induce depolarization of the cell membrane, and by forming a hole, it causes the loss of intracellular contents, thereby exhibiting antifungal action. Caspofungin, an antifungal agent of the Echinocandins family, has the effect of reversibly inhibiting the formation of the fungal cell wall, and is different from the antifungal agents that act on the cell membranes mentioned above in that it acts on the cell wall. Since azole-based drugs may cause death due to hepatitis when used in patients with reduced liver function, liver function tests must be performed prior to administration. It has been reported that flucytosine dose-dependently exhibits bone marrow suppression and hepatotoxicity, and may cause small bowel colitis, and these side effects are further increased when renal function is degraded, so it is very important to monitor the renal function of the patient. It is also prohibited for use by pregnant women. The typical toxicity of amphotericin B is glomerular nephrotoxicity due to renal artery constriction. Since it is dose-dependent, the incidence of permanent renal function loss increases when the lifetime cumulative dose is 4-5 g. In addition, nephrotoxicity such as excessive loss of potassium, magnesium, and bicarbonate due to tubular toxicity and decreased production of hematopoietic hormone may occur. Other acute reactions may include thrombophlebitis, chills, tremors, and hyperventilation.
한편, 크립토코커스 네오포만스(Cryptococcus neoformans)는 면역력이 약화된 집단에서 수막뇌염을 유발하는 담자균 균질 병원균이고 전 세계적으로 매년 60만명 이상이 사망하는 원인이다(비특허문헌 1). 하지만, 크립토코커스증(cryptococcosis) 치료에는 제한된 치료 옵션만이 가능한 상태이다(비특허문헌 2). 따라서, 크립토코커스의 다양한 생물학적 특징에 대한 완전한 이해를 통해 신규 치료 표적 및 치료 방법의 개발에 필요한 실정이다. 지난 수십년 동안, 상기 목적을 달성하기 위해 크립토코커스 네오포만스의 일반적인 생물학적 특징과 병원성을 조절하는 신호전달 캐스케이드가 광범위하게 연구되고 있다. 이러한 연구는 크립토코커스 네오포만스 내 Ras, cAMP/단백질 키나아제 A, Rim101, 칼모듈린/ 칼시뉴린(calmodulin/calcineurin), 3가지 MAPK(Cek1, Mpk1, Hog1), UPR(Unfolded Protein Response), 및 철/구리 감지를 포함하는 다양한 주요 대사 경로 및 신호전달 경로에 대한 이해를 가능하게 하였다(비특허문헌 3, 비특허문헌 4, 및 비특허문헌 5). 본 발명자들은 크립토코커스 네오포만스에서 새롭게 동정한 Ire1 및 Hxl1(HAC1 및 XBP1-유사 유전자) 단백질 및 이를 코딩하는 유전자의 기능을 손상시키는 경우, 항균 또는 뇌수막염 치료 효과가 있음을 검증하였다(특허문헌1, 특허문헌 2). 기존에 알려진 대부분의 신호전달 캐스케이드는 센서/수용체-유사 단백질 및 키나아제/포스파타아제로 구성되어 있고, 독특한 어댑터 단백질 또는 스캐폴드 단백질을 포함하고 있어서 각각의 신호전달 경로의 특이성을 향상시키며 신호전달 경로 사이의 비정상적인 혼선을 예방하였다. 그럼에도 불구하고, 각각의 신호전달 캐스케이드는 궁극적으로 하나의 전사 인자(TF, Transcription Factor) 또는 다수의 전사 인자를 활성화 또는 억제시키며, 그렇게 함으로써 전사인자의 전사 수준을 조절하는 표적 유전자 내 프로모터의 특별한 영역에 대한 전사인자의 결합에 의해 전사인자의 효과기 단백질을 증가-조절 또는 감소-조절하였다. 따라서, 전사인자는 주어진 신호전달 경로 내 유전자 발현의 주된 조절인자로 인식되고 있다. 특히, 전사인자 목록은 종 간에 다른 신호전달 구성 요소보다 보다 세분화되어 있다. 최근의 유전체 분석 결과에 기재된 바와 같이, 크립토코커스 네오포만스의 경우에는 특별히 전사인자가 더 세분화되어 있다(비특허문헌 6). 예를 들어, 소포체(ER) 스트레스 반응 및 크립토코커스 네오포만스의 독성에 중요한 UPR 신호전달 경로는 진화적으로 고도로 보존된 Ire1 키나아제 및 하류의 Hxl1 전사인자로 구성되어 있다. 따라서, 크립토코커스 네오포만스는 다양한 종류의 전사인자를 특징으로 하는 다수의 진화적으로 보존된 신호전달 캐스케이드를 갖고 있는 것처럼 보이는데, 상기 전사인자는 다른 균류의 신호전달 캐스케이드와 비교하여 독특한 크립토코커스 네오포만스의 특징을 조절할 수 있다.Meanwhile, Cryptococcus Neoformans neoformans ) is a basidiomycete homogeneous pathogen that causes meningoencephalitis in a population with weakened immunity and is the cause of more than 600,000 deaths annually worldwide (Non-Patent Document 1). However, only limited treatment options are available for the treatment of cryptococcosis (Non-Patent Document 2). Therefore, there is a need for the development of new therapeutic targets and treatment methods through a complete understanding of various biological characteristics of Cryptococcus. In the past decades, a signaling cascade that modulates the general biological characteristics and pathogenicity of Cryptococcus neoformans has been extensively studied to achieve this goal. These studies included Ras in Cryptococcus neoformans, cAMP/protein kinase A, Rim101, calmodulin/calcineurin, three MAPKs (Cek1, Mpk1, Hog1), UPR (Unfolded Protein Response), and It was possible to understand various major metabolic pathways and signaling pathways including iron/copper detection (Non-Patent
균주 전체 수준에서 크립토코커스 네오포만스 전사인자 네트워크를 이해하기 위해서, 본 발명자들은 유전체의 서열이 분석된 생명체 내 서열-특이적 DNA-결합 전사인자를 확인하기 위한 DNA-결합 도메인(DBD) 전사인자 데이터베이스를 사용하여 기존에 예측된 크립토코커스 네오포만스 내 155가지 후보 전사인자에 대하여, 상동 재조합 방법에 의해 고품질의 유전자-결실 변이체를 구축하였다(비특허문헌 7 및 비특허문헌 8). 제조된 전사인자 녹아웃(TFKO) 균주들은 30가지의 다른 생체 외 형질에 대하여 분석하였는데, 상기 형질은 성장, 분화, 스트레스 반응성, 항진균 내성, 및 독성 인자 생산에 관한 것이다. 더욱이, 본 발명에서 곤충 숙주 모델 및 쥐과 동물 숙주 모델 내 서명-표지 변이(STM, Signature-Tag Mutagenesis) 수치화를 이용하여 대규모 독성 테스트를 실시하였으며, 이에 의해 크립토코커스 네오포만스 균주 내 다양한 전사인자의 결실에 따른 형질을 분석하여 전사인자에 대한 표현형 데이터세트(발형체, phenome)을 구축함으로써 본 발명을 완성하였다.In order to understand the Cryptococcus neoformans transcription factor network at the whole strain level, the present inventors have DNA-binding domain (DBD) transcription factors for identifying sequence-specific DNA-binding transcription factors in organisms whose genome sequences are analyzed. For 155 candidate transcription factors in Cryptococcus neoformans previously predicted using the database, high-quality gene-deleting variants were constructed by homologous recombination method (Non-Patent
또한, 본 발명에 따른 크립토코커스 네오포만스 전사인자에 대한 발형체(phenome)는 온라인 (http://tf.cryptococcus.org)에서 손쉽게 접속할 수 있고, 크립토코커스 네오포만스의 일반적인 생물학적 특징을 이해할 수 있도록 도울 뿐만 아니라 크립토코커스증의 치료에 필요한 신규 표적을 제공하였다.In addition, the phenome for the Cryptococcus neoformans transcription factor according to the present invention can be easily accessed online (http://tf.cryptococcus.org), and the general biological characteristics of Cryptococcus neoformans can be understood. In addition to helping to provide a new target for the treatment of cryptococcus.
이와 같이, 본 발명은 크립토코커스 네오포만스 균주 내 전사인자 네트워크를 구축하여 상기 전사인자의 다양한 용도를 제공하는 것을 목적으로 한다.As described above, an object of the present invention is to provide various uses of the transcription factors by constructing a transcription factor network in a Cryptococcus neoformans strain.
본 발명은 크립토코커스 네오포만스 균주에 의한 진균 감염 예방 또는 치료용 항균제의 스크리닝 방법 또는 뇌수막염 치료제 스크리닝 방법을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a screening method for an antibacterial agent for preventing or treating a fungal infection caused by a Cryptococcus neoformans strain or a screening method for treating meningitis.
또한, 본 발명은 기존의 항균제 또는 뇌수막염 치료제와 병용투여시 상승효과를 가질 수 있는 후보물질의 스크리닝 방법을 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a screening method for candidate substances that can have a synergistic effect when co-administered with an existing antibacterial agent or a meningitis therapeutic agent.
본 발명의 다른 목적은, 크립토코커스 네오포만스 균주의 독성을 조절하는 전사인자 및 이를 암호화하는 유전자의 발현을 증가 또는 억제하여 항균 또는 뇌수막염 치료 효과가 있는 약학조성물을 제공하는 것을 목적으로 한다.Another object of the present invention is to provide a pharmaceutical composition having an antibacterial or meningitis treatment effect by increasing or suppressing the expression of a transcription factor that regulates the toxicity of a Cryptococcus neoformans strain and a gene encoding the same.
본 발명의 또 다른 목적은, 크립토코커스 네오포만스 균주의 항균제 감수성을 조절하는 전사인자, 성장을 조절하는 전자인자, 교배를 조절하는 전사인자, 다양한 외부 스트레스에 대한 반응을 조절하는 전사인자 및 상기 전사인자를 암호화하는 유전자의 발현을 증가 또는 억제하여 항균 또는 뇌수막염 치료 효과가 있는 약학 조성물을 제공하는 것을 목적으로 한다.Another object of the present invention is a transcription factor that regulates antimicrobial sensitivity of Cryptococcus neoformans strain, an electron factor that regulates growth, a transcription factor that regulates mating, a transcription factor that regulates the response to various external stresses, and the above An object of the present invention is to provide a pharmaceutical composition having an antibacterial or meningitis treatment effect by increasing or suppressing the expression of a gene encoding a transcription factor.
상기 목적을 달성하기 위하여, 본 발명자들은 기탁번호 KCCM51291로 기탁된 크립토코커스 네오포만스 균주를 제공한다. In order to achieve the above object, the present inventors provide a Cryptococcus neoformans strain deposited under the accession number KCCM51291.
또한, 본 발명자들은 크립토코커스 네오포만스 내 독성 조절 유전자 발현의 증가 또는 감소를 측정하는 것을 특징으로 하는 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제의 스크리닝 방법을 제공한다.In addition, the present inventors provide a screening method for an antimicrobial agent, an antimicrobial agent for co-administration, or a meningitis therapeutic agent, characterized in that the increase or decrease of the expression of a toxicity regulating gene in Cryptococcus neoformans is measured.
일 구체예에서, (a) 독성 조절 유전자를 포함하는 세포에 분석할 시료를 접촉시키는 단계; (b) 상기 세포에서 독성 조절 유전자의 발현을 측정하는 단계; (c) 상기 단계에서 독성 조절 유전자의 발현이 감소조절(down-regulation) 또는 증가조절(up-regulation)되는 것으로 측정될 때, 상기 시료가 항균제임을 판별하는 단계를 포함하는 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제 스크리닝 방법을 제공한다.In one embodiment, (a) contacting a sample to be analyzed with cells containing a toxicity regulating gene; (b) measuring the expression of a toxicity regulatory gene in the cell; (c) When the expression of the toxicity regulating gene is determined to be down-regulated or up-regulated in the above step, an antimicrobial agent comprising the step of determining that the sample is an antimicrobial agent, an antimicrobial agent for co-administration Or it provides a meningitis treatment screening method.
상기 구체예에서, 상기 독성 조절 유전자는 크립토코커스 네오포만스의 병원성을 조절하는 유전자로서 발현이 감소되는 유전자는 usv101, fzc1, bap1, hob1, zfc2, fzc50, fzc31, bzp2, fzc9, ddt1, mal13, fzc2, fzc43, fzc22, hlh1, mbs2, rum1, fzc5, aro80, clr1, pip2, fzc37, gat5, fzc49, cef3, fzc33, fzc12 및 zfc5를 포함하는 군으로부터 선택된 어느 하나의 유전자이고, 발현이 증가하는 유전자는 fzc17, fzc40, aro8001, fzc38, fzc24 및 ert1을 포함하는 군으로부터 선택된 어느 하나의 유전자인 항균제, 병용투영용 항균제 또는 뇌수막염 치료제의 스크리닝 방법을 제공한다.In the above embodiment, the toxicity regulating gene is a gene that regulates the pathogenicity of Cryptococcus neoformans, and the gene whose expression is reduced is usv101 , fzc1 , bap1 , hob1 , zfc2 , fzc50 , fzc31 , bzp2 , fzc9 , ddt1 , mal13 , fzc2 , fzc43 , fzc22 , hlh1 , mbs2 , rum1 , fzc5 , aro80 , clr1 , pip2 , fzc37 , gat5 , fzc49 , cef3 , fzc33 , fzc12, and zfc5 . Provides a screening method of an antibacterial agent, an antibacterial agent for co-projection, or a treatment for meningitis, which is any one gene selected from the group comprising fzc17 , fzc40 , aro8001 , fzc38 , fzc24, and ert1.
상기 구체예에서, 상기 독성 조절 유전자는 캡슐, 멜라닌 및 우레아제를 포함하는 군으로부터 선택된 어느 하나의 생산을 조절하는 것인 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제의 스크리닝 방법을 제공한다.In the above embodiment, the toxicity regulating gene provides a screening method for an antimicrobial agent, an antimicrobial agent for co-administration, or a meningitis therapeutic agent that regulates the production of any one selected from the group comprising capsules, melanin, and urease.
상기 구체예에서, 상기 캡슐 생산을 감소시키는 유전자는 bap1, rds2, zap104, fzc47, gat204, fzc33, fzc45, hsf2, bzp4, hob5, fzc16, hob3, zfc4, mcm1, liv4, hob4 및 liv1을 포함하는 군으로부터 선택된 어느 하나의 유전자이고 상기 캡슐 생산을 증가시키는 유전자는 hob7, clr3, fzc51, fzc1, fkh2, nrg1, usv101, fzc29, bzp3, zfc3, fzc14, sre1, fzc30, hlh4, fzc36, crl6, mln1, fzc46, clr1, fzc17, jjj1, fzc49, fzc18, hcm1, fzc24, hlh3 및 hpa1을 포함하는 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제의 스크리닝 방법을 제공한다.In the above embodiment, the gene for reducing capsule production is a group comprising bap1 , rds2 , zap104 , fzc47 , gat204 , fzc33 , fzc45 , hsf2 , bzp4 , hob5 , fzc16 , hob3 , zfc4 , mcm1 , liv4 , hob4, and liv1 . Any one gene selected from and the gene that increases the capsule production is hob7 , clr3 , fzc51 , fzc1 , fkh2 , nrg1 , usv101 , fzc29 , bzp3 , zfc3 , fzc14 , sre1 , fzc30 , hlh4 , fzc36 , crl6 , mln1 , fzc46 , clr1 , fzc17 , jjj1 , fzc49 , fzc18 , hcm1 , fzc24 , hlh3 and hpa1 It provides a screening method of an antibacterial agent, an antibacterial agent for co-administration, or a meningitis therapeutic agent, characterized in that it is any one gene selected from the group consisting of.
상기 구체예에서, 상기 멜라닌 생산을 감소시키는 유전자는 bzp4, fzc8, hob1, usv101, liv1, mbs2, fzc5, fzc25 및 ert1을 포함하는 군으로부터 선택된 어느 하나의 유전자이고 상기 멜라닌 생산을 증가시키는 유전자는 bzp2, fkh2, bap1, bzp3, hlh1, sip4, rds2, sip401, fzc1, gat1, ada2, nrg1, fzc31 및 hlh2를 포함하는 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제의 스크리닝 방법을 제공한다.In the above embodiment, the gene for reducing melanin production is any one gene selected from the group comprising bzp4 , fzc8 , hob1 , usv101 , liv1 , mbs2 , fzc5 , fzc25, and ert1 , and the gene that increases melanin production is bzp2. , fkh2 , bap1 , bzp3 , hlh1 , sip4 , rds2 , sip401 , fzc1 , gat1 , ada2 , nrg1 , fzc31 and hlh2 . Provides a screening method of.
상기 구체예에서, 상기 우레아제 생산을 감소시키는 유전자는 zap104, sre1, gat201, fzc46, hlh1 및 fzc21을 포함하는 군으로부터 선택된 어느 하나의 유전자이고 상기 우레아제 생산을 증가시키는 유전자는 rim1, atf1, fkh2, fzc1, usv101, bap1, sxi1alpha, mln1, fzc26, skn7, zfc7, hob7, fzc14 및 hob4를 포함하는 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제의 스크리닝 방법을 제공한다.In the above embodiment, the gene for reducing the urease production is any one gene selected from the group comprising zap104 , sre1 , gat201 , fzc46 , hlh1 and fzc21 , and the gene for increasing the urease production is rim1 , atf1 , fkh2 , fzc1 , usv101 , bap1 , sxi1alpha , mln1 , fzc26 , skn7 , zfc7 , hob7 , fzc14, and hob4 . Provides a screening method for an antibacterial agent, an antimicrobial agent for combined administration, or a meningitis therapeutic agent, characterized in that it is any one gene selected from the group comprising .
상기 구체예에서, 상기 세포는 크립토코커스 네오포만스인 것을 특징으로 하는 항균제, 병용 투여용 항균제 또는 뇌수막염 치료제 스크리닝 방법을 제공한다.In the above embodiment, the cell is a Cryptococcus neoformans, it provides an antibacterial agent, an antimicrobial agent for co-administration, or a meningitis therapeutic agent screening method.
본 발명의 스크리닝 방법을 언급하면서 사용되는 용어 "시료"는 유전자의 발현량에 영향을 미치거나, 단백질의 양 또는 활성에 영향을 미치는지 여부를 검사하기 위하여 스크리닝에서 이용되는 미지의 후보 물질을 의미한다. 상기 시료는 화학물질, 뉴클레오타이드, 안티센스-RNA, siRNA(small interference RNA) 및 천연물 추출물을 포함하나, 이에 한정되는 것은 아니다.The term "sample" used while referring to the screening method of the present invention refers to an unknown candidate substance used in screening to examine whether it affects the expression level of a gene or affects the amount or activity of a protein. . The sample includes, but is not limited to, chemicals, nucleotides, antisense-RNA, small interference RNA (siRNA), and natural product extracts.
본 발명에서 "항균제"는 세균 및/또는 곰팡이류의 번식을 억제하는 것으로, 무기계 항균제, 유기계 천연물 추출계 항균제, 유기계 지방족 화합물 항균제 및 유기계 방향족 화합물 항균제를 포함한다. 또한, 이에 한정되지 않지만, 무기계 항균제로서는, 차아염소 나트륨으로 대표되는 염소 화합물, 과산화 수소로 대표되는 과산화물 붕산, 붕산 나트륨으로 대표되는 붕산 화합물, 황산동으로 대표되는 동화합물 황산 아연, 염화 아연으로 대표되는 아연 화합물 유황, 다황산 석회, 수화 유황으로 대표되는 유황계물 산화 칼슘으로 대표되는 칼슘 화합물 티오술파이트 은착염, 질산은으로 대표되는 은화합물, 그 밖에, 옥소, 실리코플루오리드 나트륨 등을 들 수 있고, 유기계 천연물 추출계 항균제로서는, 히노키티올, 맹종죽 추출액, 크레오소트유 등을 들 수 있다.In the present invention, the "antibacterial agent" is to inhibit the propagation of bacteria and/or fungi, and includes an inorganic antibacterial agent, an organic natural product extracting antibacterial agent, an organic aliphatic compound antibacterial agent, and an organic aromatic compound antibacterial agent. In addition, although not limited thereto, examples of the inorganic antibacterial agent include a chlorine compound represented by sodium hypochlorite, a peroxide boric acid represented by hydrogen peroxide, a boric acid compound represented by sodium borate, an anabolic compound represented by copper sulfate, zinc sulfate, and zinc chloride. Zinc compound sulfur, lime polysulfate, calcium compound represented by sulfur-based calcium oxide represented by hydrated sulfur, thiosulfite silver complex salt, silver compound represented by silver nitrate, and in addition, oxo, sodium silicofluoride, and the like. Examples of the organic-based natural product extract-based antibacterial agent include hinokitiol, a maengjongguk extract, creosote oil, and the like.
본 발명에서 "뇌수막염"은 거미막과 연질막 사이에 존재하는 거미막밀 공간(subarachnoid space)에 염증이 발생하는 다양한 질환으로, 거미막밀공간에 바이러스나 세균이 침투하여 발생하는 수막염이지만, 특정 화학 물질에 의한 염증, 암세포의 뇌척수액 공간으로의 파종에 의해 발생하는 것을 말한다.In the present invention, "meningitis" is a variety of diseases in which inflammation occurs in the subarachnoid space existing between the arachnoid and soft membranes, and is meningitis caused by the penetration of viruses or bacteria into the arachnoid space, but specific chemical substances It is caused by inflammation caused by and the seeding of cancer cells into the cerebrospinal fluid space.
본 발명은 크립토코커스 네오포만스 균주의 독성을 조절하는 전사 인자의 발현 수준을 탐지하여 항균 효과 또는 뇌수막염 치료효과를 가지는 조성물을 효과적으로 스크리닝할 수 있다. 또한, 크립토코커스 네오포만스의 독성을 조절하는 전사 인자의 발현을 증가 또는 감소 조절함으로써 항균 효과 또는 뇌수막염 치료효과를 나타내는 약학 조성물을 제공할 수 있다.The present invention can effectively screen a composition having an antibacterial effect or a meningitis treatment effect by detecting the expression level of a transcription factor that regulates the toxicity of the Cryptococcus neoformans strain. In addition, it is possible to provide a pharmaceutical composition exhibiting an antibacterial effect or a meningitis treatment effect by increasing or decreasing the expression of a transcription factor that regulates the toxicity of Cryptococcus neoformans.
도 1a 내지 도 1h는 본 발명에 따른 크립토코커스 네오포만스 전사인자들의 블라스트 매트릭스를 나타낸 것이다.
도 2a 내지 도 2y는 본 발명에 따른 크립토코커스 네오포만스 전사인자들의 블라스트 분석 결과를 나타낸 것이다.
도 3a 및 도3b는 크립토코커스 네오포만스 전사인자의 개요, 전반적인 결실 및 발형체(phenome)-기반 분석에 대한 전략에 관한 것으로, 도 3a는 크립토코커스 네오포만스 전사인자의 종류 및 분류를 나타내는 파이 그래프(pie chart)이고, 도 3b는 크립토코커스 네오포만스 전사인자 녹아웃(TFKO) 라이브러리 구축, 발형체-기반 분석에 대한 순서도(flow chart)를 나타낸 것이다.
도 4a 내지 도 4r은 크립토코커스 네오포만스 전사인자 녹아웃 균주에 대하여 제작된 발형체 열 지도(heat map)을 나타낸 것인데, 도 4a는 전체 결과를 나타낸 것이고 도 4b 내지 도 4r은 도 4a를 세분한 결과를 나타낸 것이다.
도 5a 및 도 5b는 크립토코커스 네오포만스의 온도-의존적 성장에 필요한 전사 인자를 나타낸 것이다.
도 6a 내지 도 6d는 크립토코커스 네오포만스의 성 분화에 관련된 전사 인자를 나타낸 것으로, 도 6a는 교배 실험에서 야생형 균주 H99 및 각각의 전사인자 변이체를 V8 배지 상의 대립 교배 형 KN99a 균주와 공통배양하여 빛을 차단하고 7일간 상온에서 배양한 결과를 나타낸 것이고, 도 6b는 각각의 전사인자 변이체의 세포-융합 효율에 관한 것으로 대조군 균주의 효율과 비교하여 계산한 결과를 나타낸 것이며[NAT-표시된 야생형균주 YSB119 NEO-표시된 야생형 a 균주 YSB121], 도 6c는 페로몬 유전자 발현에 관련된 전사 인자를 나타낸 것으로 표시된 전사인자 변이체는 KN99a 균주와 V8 배지 상에 상온에서 18 내지 24시간 동안 공통배양한 후 RNA 발현을 분석한 결과를 나타낸 것이며, 도 6d는 크립토코커스 네오포만스 균주의 교배 단계에서 표시된 전사인자의 역할을 나타낸 모식도이다.
도 7a 및 도 7b는 크립토코커스 네오포만스 내 교배 효율(성 분화)에 관련되어 있는 전사인자를 나타낸 것이다(7a-양성조절인자, 7b-음성조절인자).
도 8a 내지 도 8e는 크립토코커스 네오포만스 내 독성-인자 생산에 관련된 전사인자를 나타낸 것으로, 도 8a는 야생형 균주 H99 및 전사인자 변이체에서 생산되는 캡슐의 크기를 나타낸 것이고, 도 8b 및 도 8c는 크립토코커스 네오포만스 내 캡슐 생산에 관련된 전사인자(8b-음성조절인자, 8c-양성조절인자)를 나타낸 것이며, 도 8d 및 도 8e는 멜라닌 합성에 관련된 주요 락카제(laccase)인 LAC1 발현과 전사인자 사이의 관련성에 관한 것으로 도 8d는 전사인자 변이체를 니거 시드 아가 배지(0.1% 글루코스 및 0.3% 글루코스 포함) 상에 스팟팅하여 37℃에서 배양하면서 촬영한 결과를 나타낸 것이며, 도 8e는 글루코스-풍족 조건(0h) 및 글루코스-결핍 조건(1h 및 2h)의 세포로부터 추출한 전체 RNA에 대하여 LAC1-특이 탐침을 이용한 노던 블롯 분석 결과를 나타낸 것이다.
도 9a 및 도 9b는 크립토코커스 네오포만스 내 멜라닌 생산에 관련되어 있는 전사인자를 나타낸 것이다(9a-양성조절인자, 9b-음성조절인자).
도 10a 및 도 10b는 크립토코커스 네오포만스 내 우레아제 생산에 필요한 전사인자를 나타낸 것이다(10a-음성조절인자, 10b-양성조절인자).
도 11a 내지 도 11d는 벌집나방(Galleria mellonella) 살상 실험에 의해 크립토코커스 네오포만스의 독성에 관련된 전사인자를 나타낸 것으로, 도 11a 내지 도 11c는 여러 가지 전자인자 결실체에 대한 결과를 나타낸 것이고 도 11d는 서명-표지 돌연변이 유발(STM)-기반의 마우스 실험에 의해 크립토코커스 내 독성에 관련된 전사인자를 검증한 결과를 나타낸 것이다.
도 12a 내지 도 12d는 전사인자 변이체의 STM 수치를 나타낸 것이다.
도 13은 크립토코커스 네오포만스 내 항균제 내성에 관련된 전사인자를 나타낸 것이다.
도 14a 내지 도 14e는 크립토코커스 네오포만스 내 스테롤-생합성 유전자를 조절하는 전사인자들이 일반적인 환경 스트레스 반응 및 적응을 조절한다는 것을 나타내는 결과인데, 도 14a는 8가지 전사인자 변이체의 항균제에 대한 반응을 관찰한 결과이고, 도 14b는 여러 가지 전사인자 변이체의 플루코나졸(FCZ) 에 대하여 ERG11-특이적인 탐침을 이용한 노던 블롯 결과를 나타낸 것이며, 도 14c는 sre1 변이체 및 hob1 변이체에 플루코나졸(FCZ)에 대하여 ERG 유전자 특이적인 탐침을 이용한 노던 블롯 결과를 나타낸 것이며, 도 14d는 sre1 변이체 및 hob1 변이체의 다양한 스트레스-유도 물질에 대한 반응을 관찰한 결과이며, 도 14e는 크립토코커스 네오포만스 내 스테롤 항상성 및 일반적인 스트레스 반응성에서 Sre1 및 Hob1의 역할에 대하여 제시된 모델을 나타낸 것이다.
도 15a 내지 도 15f는 다양한 스트레스에 대한 크립토코커스 네오포만스의 스트레스 반응 및 적응에 관련된 전사인자를 나타낸 것이다(a.삼투압 스트레스; b. 산화 스트레스; c. 소포체 스트레스; d. 유전독성 스트레스; e. 세포벽 및 세포막 스트레스; f. 중금속 스트레스).
도 16은 병원성과 발형체 사이의 기능성 상관관계를 나타낸 것이다.1A to 1H show a blast matrix of Cryptococcus neoformans transcription factors according to the present invention.
2A to 2Y show the results of blast analysis of Cryptococcus neoformans transcription factors according to the present invention.
3A and 3B are schematics of Cryptococcus neoformans transcription factors, general deletions, and strategies for phenome-based analysis, and FIG. 3A shows the types and classifications of Cryptococcus neoformans transcription factors. It is a pie chart, and FIG. 3B shows a flow chart for Cryptococcus neoformans transcription factor knockout (TFKO) library construction and morphogen-based analysis.
Figures 4a to 4r show the developmental body heat map (heat map) produced for the Cryptococcus neoformans transcription factor knockout strain, Figure 4a shows the overall results and Figures 4b to 4r are subdivided into Figure 4a It shows the results.
5A and 5B show transcription factors required for temperature-dependent growth of Cryptococcus neoformans.
6A to 6D show transcription factors related to sexual differentiation of Cryptococcus neoformans, and FIG. 6A is co-cultured with wild-type strain H99 and each of the transcription factor variants in a crossing experiment with allelic KN99a strains on V8 medium. It shows the result of culturing at room temperature for 7 days after blocking the light, and Fig. 6b shows the result calculated by comparing the efficiency of the control strain as related to the cell-fusion efficiency of each transcription factor variant [NAT-indicated wild-type strain YSB119 NEO-labeled wild type a strain YSB121], Figure 6c is a transcription factor variant indicated to show a transcription factor related to pheromone gene expression after co-culture for 18 to 24 hours at room temperature on KN99a strain and V8 medium, and then analyzed RNA expression One result is shown, and FIG. 6D is a schematic diagram showing the role of the transcription factor indicated in the mating stage of the Cryptococcus neoformans strain.
7A and 7B show transcription factors involved in mating efficiency (sex differentiation) in Cryptococcus neoformans (7a-positive regulator, 7b-negative regulator).
Figures 8a to 8e show the transcription factors involved in the production of toxicity-factors in Cryptococcus neoformans, Figure 8a shows the size of the capsules produced in the wild-type strain H99 and the transcription factor variant, Figures 8b and 8c It shows the transcription factors (8b-negative regulator, 8c-positive regulator) related to capsule production in Cryptococcus neoformans, and Figures 8D and 8E are LAC1 expression and transcription, a major laccase related to melanin synthesis. Regarding the relationship between the factors, FIG. 8D shows the results obtained while culturing at 37° C. by spotting the transcription factor variants on Niger's seed agar medium (including 0.1% glucose and 0.3% glucose), and FIG. 8E is glucose- The results of Northern blot analysis using a LAC1 -specific probe are shown for total RNA extracted from cells under abundance conditions (0h) and glucose-deficient conditions (1h and 2h).
9A and 9B show transcription factors involved in melanin production in Cryptococcus neoformans (9a-positive regulator, 9b-negative regulator).
10A and 10B show transcription factors required for urease production in Cryptococcus neoformans (10a-negative regulator, 10b-positive regulator).
11A to 11D show transcription factors related to the toxicity of Cryptococcus neoformans by a honeycomb moth (Galleria mellonella) killing experiment, and FIGS. 11A to 11C show results for various electron factor deletion bodies. 11d shows the results of verifying transcription factors related to toxicity in Cryptococcus by a signature-labeled mutagenesis (STM)-based mouse experiment.
12A to 12D show the STM values of the transcription factor variants.
13 shows transcription factors related to antimicrobial resistance in Cryptococcus neoformans.
14A to 14E are results showing that transcription factors regulating sterol-biosynthetic genes in Cryptococcus neoformans regulate general environmental stress response and adaptation, and FIG. 14A shows the response of eight transcription factor variants to antimicrobial agents. Figure 14b shows the Northern blot results using an ERG11-specific probe for fluconazole (FCZ) of various transcription factor variants, and Figure 14c shows ERG for fluconazole (FCZ) in the sre1 variant and the hob1 variant. It shows the Northern blot result using a gene-specific probe, and FIG. 14D is a result of observing the response of the sre1 mutant and the hob1 mutant to various stress-inducing substances, and FIG. The proposed model for the role of Sre1 and Hob1 in reactivity is shown.
15A to 15F show transcription factors related to the stress response and adaptation of Cryptococcus neoformans to various stresses (a. osmotic stress; b. oxidative stress; c. endoplasmic reticulum stress; d. genotoxic stress; e) Cell wall and cell membrane stress; f. heavy metal stress).
Figure 16 shows the functional correlation between pathogenicity and proforma.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당 업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시예Example
전사인자 Transcription factor 변이체(TF mutant)의Of TF mutant 제작 making
1.1 전사인자 1.1 transcription factor 변이체Variant 대상 전사인자 선별 Selection of target transcription factors
공개된 DBD 전사인자(TF, Transcription Factor) 예측 데이터베이스 (http://transcriptionfactor.org/) (비특허문헌 8)를 사용하여 추정되는 전사인자를 선별하였다. 크립토코커스 네오포만스 H99 균주(혈청형 A 게놈-서열 플랫폼 균주)는 188가지 전사 인자(Pfam으로부터 예측되는 148 가지 및 수퍼패밀리로부터 예측되는 96가지)를 포함한다. 이러한 전사인자는 H99 유전체 데이터베이스 1차 버전에 근거하여 예측되므로, 본 발명자는 H99 유전체 데이터베이스(비특허문헌 6)의 가장 최신 버전(버전 7)에 대한 자료를 이용하여 H99 유전체 데이터베이스에 대한 업데이트를 실시하였다. 업데이트 결과에 따라 최종적으로 155가지 전사인자를 선별하였다(표 1). BLAST e-수치 매트릭스에 근거한 이종상동성 매핑 결과는 크립토코커스 네오포만스 균주가 진화적으로 구별되는 다수 그룹의 전사인자를 포함하는 것을 개시하고 있다(도 1). 크립토코커스 DNA 결합 도메인(DNA Binding Domain) 전사인자는 DNA 결합 도메인(DBD, DNA Binding Domain)에 근거하여 분류하였다(도 1a). 상기 전사인자의 44%(78가지)는 진균 Zn2-Cys6 DBD를 포함하였고, 이 중에서 40가지는 진균-특이적인 도메인을 포함하고 있다. 여러 가지 전사인자는 2가지 이상의 전사인자 도메인을 포함하고 있다(표 1). The estimated transcription factor was selected using the published DBD transcription factor (TF, Transcription Factor) prediction database (http://transcriptionfactor.org/) (Non-Patent Document 8). The Cryptococcus neoformans H99 strain (serum type A genome-sequence platform strain) contains 188 transcription factors (148 predicted from Pfam and 96 predicted from the superfamily). Since these transcription factors are predicted based on the first version of the H99 genome database, the present inventors update the H99 genome database using the data on the most recent version (version 7) of the H99 genome database (non-patent document 6). I did. Finally, 155 transcription factors were selected according to the update results (Table 1). The results of orthologous mapping based on the BLAST e-value matrix disclose that the Cryptococcus neoformans strains contain evolutionarily distinct groups of transcription factors (FIG. 1). Cryptococcus DNA binding domain (DNA Binding Domain) transcription factor was classified based on the DNA binding domain (DBD, DNA Binding Domain) (Fig. 1a). 44% (78) of the transcription factors included the fungal Zn2-Cys6 DBD, of which 40 contained fungi-specific domains. Various transcription factors contain two or more transcription factor domains (Table 1).
(Gene name)Gene name
(Gene name)
1
One
02566
02566
"Winged helix" DNA-binding domain / Fork head domain
"Winged helix" DNA-binding domain / Fork head domain
FKH2FKH2
2
2
00791
00791
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
HLH1HLH1
3
3
01069
01069
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC11FZC11
4
4
07464
07464
APSES domain / Ankyrin repeat#2
APSES domain /
MBS1MBS1
5
5
03401
03401
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT203GAT203
6
6
04588
04588
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
ERT1ERT1
7
7
00828
00828
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
SIP401SIP401
8
8
03561
03561
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC33FZC33
9
9
06762
06762
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT204GAT204
10
10
06276
06276
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
CEP3CEP3
11
11
05785
05785
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
STB4STB4
12
12
03132
03132
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC5FZC5
13
13
01438
01438
KilA-N domain / Ankyrin repeats (many copies)
KilA-N domain / Ankyrin repeats (many copies)
MBS2MBS2
14
14
07593
07593
bZIP transcription factor / Domain of unknown function (DUF3425)
bZIP transcription factor / Domain of unknown function (DUF3425)
YAP4YAP4
15
15
04837
04837
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
MLN1MLN1
16
16
05093
05093
Homeobox domain
Homeobox domain
HOB6HOB6
17
17
05642
05642
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC37FZC37
18
18
05431
05431
Zinc-finger double domain / C2H2-type zinc finger
Zinc-finger double domain / C2H2-type zinc finger
RIM101RIM101
19
19
04398
04398
Fungal specific transcription factor domain
/ Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal specific transcription factor domain
/ Fungal Zn(2)-Cys(6) binuclear cluster domain
ARO80ARO80
20
20
04878
04878
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC1FZC1
21
21
03183
03183
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC24FZC24
22
22
03710
03710
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
ECM22ECM22
23
23
03279
03279
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
CCD4CCD4
24
24
04637
04637
Helix-turn-helix/lambda repressor-like DNA-binding domains
Helix-turn-helix/lambda repressor-like DNA-binding domains
MBF1MBF1
25
25
06425
06425
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
PPR1PPR1
26
26
04345
04345
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
ARO8001ARO8001
27
27
04184
04184
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC47FZC47
28
28
02774
02774
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
MAL13MAL13
29
29
00670
00670
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC12FZC12
30
30
00068
00068
Zinc-finger double domain / C2H2-type zinc finger
Zinc-finger double domain / C2H2-type zinc finger
MET32MET32
31
31
05010
05010
C2H2-type zinc finger
C2H2-type zinc finger
ZFC7ZFC7
32
32
04090
04090
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
ATF1ATF1
33
33
06134
06134
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
BZP1BZP1
((
HXL1HXL1
))
34
34
04630
04630
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
BAP2BAP2
35
35
07901
07901
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC29FZC29
36
36
01173
01173
Beta-trefoil DNA-binding domain / p53-like transcription factors / DNA-binding protein LAG-1 (CSL)
Beta-trefoil DNA-binding domain / p53-like transcription factors / DNA-binding protein LAG-1 (CSL)
PAN1PAN1
37
37
03115
03115
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC46FZC46
38
38
07924
07924
SRF-type transcription factor (DNA-binding and dimerisation
SRF-type transcription factor (DNA-binding and dimerisation
MCM1MCM1
39
39
07435
07435
CCAAT-binding transcription factor (CBF-B/NF-YA) subunit B
CCAAT-binding transcription factor (CBF-B/NF-YA) subunit B
HAP2HAP2
40
40
02555
02555
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
SIP402SIP402
41
41
04594
04594
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC27FZC27
42
42
06188
06188
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC15FZC15
43
43
05170
05170
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
PIP2PIP2
44
44
02241
02241
Homeodomain-like / Helix-turn-helix domain
Homeodomain-like / Helix-turn-helix domain
HOB5HOB5
45
45
06921
06921
Homeobox domain
Homeobox domain
HOB4HOB4
46
46
05186
05186
GRF zinc finger
GRF zinc finger
GRF1GRF1
47
47
00896
00896
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC34FZC34
48
48
00039
00039
C2H2-type zinc finger
C2H2-type zinc finger
ZFC6ZFC6
49
49
07940
07940
Basic region leucine zipper / bZIP transcription factor
Basic region leucine zipper / bZIP transcription factor
BZP5BZP5
50
50
06814
06814
Homeobox KN domain
Homeobox KN domain
SXI1alphaSXI1alpha
51
51
01454
01454
STE like transcription factor / Zinc-finger double domain
/ C2H2-type zinc finger
STE like transcription factor / Zinc-finger double domain
/ C2H2-type zinc finger
STE12STE12
52
52
03527
03527
C2H2-type zinc finger / C3HC4-type zinc finger
C2H2-type zinc finger / C3HC4-type zinc finger
HEL2HEL2
53
53
05255
05255
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC2FZC2
54
54
05112
05112
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC42FZC42
55
55
01883
01883
Glucocorticoid receptor-like (DNA-binding domain) / GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain) / GATA zinc finger
GAT8GAT8
56
56
04353
04353
C2H2-type zinc finger
C2H2-type zinc finger
CLR1CLR1
57
57
05375
05375
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
HLH2HLH2
58
58
03998
03998
SRF-type transcription factor (DNA-binding and dimerisation
SRF-type transcription factor (DNA-binding and dimerisation
RLM1RLM1
59
59
00239
00239
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
BAP1BAP1
60
60
06871
06871
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC41FZC41
61
61
00156
00156
C2H2-type zinc finger
C2H2-type zinc finger
SP1(SP1(
CRZ1CRZ1
))
62
62
04268
04268
GRF zinc finger
GRF zinc finger
APN2APN2
63
63
05420
05420
Zinc-finger double domain / C2H2-type zinc finger
Zinc-finger double domain / C2H2-type zinc finger
USV101USV101
64
64
00018
00018
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC6FZC6
65
65
03346
03346
bZIP transcription factor
bZIP transcription factor
BZP4BZP4
66
66
00514
00514
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT6GAT6
67
67
05538
05538
SRR1 domain / C2H2-type zinc finger / DnaJ domain
SRR1 domain / C2H2-type zinc finger / DnaJ domain
JJJ1JJJ1
68
68
03409
03409
"Winged helix" DNA-binding domain / CheY-like / HSF-type DNA-binding
"Winged helix" DNA-binding domain / CheY-like / HSF-type DNA-binding
SKN7SKN7
69
69
06339
06339
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC35FZC35
70
70
07011
07011
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC22FZC22
71
71
07506
07506
NF-X1 type zinc finger #3 / R3H domain
NF-X1 type
FAP1FAP1
72
72
04807
04807
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC8FZC8
73
73
02435
02435
PYP-like sensor domain (PAS domain)
/ Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger / AT hook motif
PYP-like sensor domain (PAS domain)
/ Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger / AT hook motif
BWC2 (CWC2)
BWC2 ( CWC2 )
74
74
02364
02364
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC19FZC19
75
75
03116
03116
"Winged helix" DNA-binding domain / Fork head domain
"Winged helix" DNA-binding domain / Fork head domain
HCM1HCM1
76
76
02877
02877
Zinc-finger double domain / Fungal Zn(2)-Cys(6) binuclear cluster domain
Zinc-finger double domain / Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC51FZC51
77
77
00559
00559
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
BZP3BZP3
78
78
03914
03914
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC14FZC14
79
79
00871
00871
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
CLR3CLR3
80
80
06483
06483
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC25FZC25
81
81
07797
07797
Putative FMN-binding domain
Putative FMN-binding domain
CRL6CRL6
82
82
05019
05019
Fungal specific transcription factor domain
Fungal specific transcription factor domain
FZC21FZC21
83
83
05380
05380
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC44FZC44
84
84
04518
04518
Fungal specific transcription factor domain / C2H2-type zinc finger
Fungal specific transcription factor domain / C2H2-type zinc finger
ZFC5ZFC5
85
85
05176
05176
Homeobox domain
Homeobox domain
HOB3HOB3
86
86
05861
05861
Fork head domain
Fork head domain
FKH101FKH101
87
87
00460
00460
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
LIV1LIV1
88
88
02305
02305
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC45FZC45
89
89
03849
03849
Fungal specific transcription factor domain
/ Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal specific transcription factor domain
/ Fungal Zn(2)-Cys(6) binuclear cluster domain
ASG1ASG1
90
90
01014
01014
C2H2-type zinc finger
C2H2-type zinc finger
ZFC4ZFC4
91
91
01858
01858
Homeobox domain
Homeobox domain
HOB2HOB2
92
92
06719
06719
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC49FZC49
93
93
04093
04093
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
YRM103YRM103
94
94
04176
04176
"Winged helix" DNA-binding domain / HSF-type DNA-binding
"Winged helix" DNA-binding domain / HSF-type DNA-binding
HSF2HSF2
95
95
01431
01431
Homeobox domain
Homeobox domain
HOB1HOB1
96
96
07443
07443
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
HLH4HLH4
97
97
04916
04916
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC16FZC16
98
98
05392
05392
Zinc-finger double domain / C2H2-type zinc finger
Zinc-finger double domain / C2H2-type zinc finger
ZAP104ZAP104
99
99
02322
02322
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC17FZC17
100
100
04012
04012
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC18FZC18
101
101
00505
00505
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC28FZC28
102
102
06751
06751
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
HLH3HLH3
103
103
04841
04841
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC43FZC43
104
104
03212
03212
"Winged helix" DNA-binding domain
"Winged helix" DNA-binding domain
HCM101HCM101
105
105
01626
01626
Zinc finger, ZZ type / Myb-like DNA-binding domain
/ SWIRM domain
Zinc finger, ZZ type / Myb-like DNA-binding domain
/ SWIRM domain
ADA2ADA2
106
106
03894
03894
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
PDR802PDR802
107
107
02066
02066
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC13FZC13
108
108
06818
06818
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
HAP1HAP1
109
109
05940
05940
C2H2-type zinc finger
C2H2-type zinc finger
ZFC3ZFC3
110
110
01948
01948
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC36FZC36
111
111
04804
04804
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
SRE1SRE1
112
112
04352
04352
Zinc-finger double domain / C2H2-type zinc finger
Zinc-finger double domain / C2H2-type zinc finger
ZAP103ZAP103
113
113
03768
03768
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC32FZC32
114
114
01977
01977
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC39FZC39
115
115
04895
04895
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC3FZC3
116
116
04583
04583
WSTF, HB1, Itc1p, MBD9 motif 2/3 / DDT domain
WSTF, HB1, Itc1p,
DDT1DDT1
117
117
01973
01973
Zinc-finger double domain / C2H2-type zinc finger
/ Fungal specific transcription factor domain
Zinc-finger double domain / C2H2-type zinc finger
/ Fungal specific transcription factor domain
ZFC2ZFC2
118
118
02603
02603
Fungal specific transcription factor domain
/ C2H2-type zinc finger
Fungal specific transcription factor domain
/ C2H2-type zinc finger
ZFC1ZFC1
119
119
04036
04036
"Winged helix" DNA-binding domain / HSF-type DNA-binding
"Winged helix" DNA-binding domain / HSF-type DNA-binding
HSF3HSF3
120
120
06156
06156
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC7FZC7
121
121
03431
03431
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC48FZC48
122
122
07922
07922
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC4FZC4
123
123
00031
00031
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
MLR1MLR1
124
124
03018
03018
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
ASG101ASG101
125
125
04263
04263
Glucocorticoid receptor-like (DNA-binding domain) / Basic region leucine zipper
/ bZIP transcription factor / GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain) / Basic region leucine zipper
/ bZIP transcription factor / GATA zinc finger
BZP2BZP2
126
126
01708
01708
GATA zinc finger
GATA zinc finger
GAT7GAT7
127
127
00332
00332
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
SIP4SIP4
128
128
07724
07724
Copper fist DNA binding domain
Copper fist DNA binding domain
CUF1CUF1
129
129
03902
03902
Fungal Zn(2)-Cys(6) binuclear cluster domain/PAS fold
Fungal Zn(2)-Cys(6) binuclear cluster domain/PAS fold
RDS2RDS2
130
130
03366
03366
Zinc-finger double domain / C2H2-type zinc finger
Zinc-finger double domain / C2H2-type zinc finger
ZNF2ZNF2
131
131
05222
05222
C2H2-type zinc finger / Zinc-finger double domain
C2H2-type zinc finger / Zinc-finger double domain
NRG1NRG1
132
132
05049
05049
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
PIP201PIP201
133
133
02516
02516
Helix-loop-helix DNA-binding domain
Helix-loop-helix DNA-binding domain
HLH5HLH5
134
134
04774
04774
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC26FZC26
135
135
03059
03059
Fungal Zn(2)-Cys(6) binuclear cluster domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC9FZC9
136
136
00193
00193
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT1GAT1
137
137
03336
03336
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC50FZC50
138
138
03086
03086
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC20FZC20
139
139
03229
03229
Homeobox domain / Homeobox KN domain
Homeobox domain / Homeobox KN domain
YOX101YOX101
140
140
01841
01841
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GLN3GLN3
141
141
02476
02476
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
YRM101YRM101
142
142
05153
05153
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger / AT hook motif
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger / AT hook motif
GAT5GAT5
143
143
02700
02700
C2H2-type zinc finger
C2H2-type zinc finger
ZFC8ZFC8
144
144
04586
04586
Homeobox domain
Homeobox domain
HOB7HOB7
145
145
02723
02723
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC23FZC23
146
146
03741
03741
Fungal Zn(2)-Cys(6) binuclear cluster domain / AT hook motif
Fungal Zn(2)-Cys(6) binuclear cluster domain / AT hook motif
FZC31FZC31
147
147
04457
04457
Fungal Zn(2)-Cys(6) binuclear cluster domain / AT hook motif
Fungal Zn(2)-Cys(6) binuclear cluster domain / AT hook motif
FZC30FZC30
148
148
06283
06283
Homeodomain-like
Homeodomain-like
LIV4LIV4
149
149
07411
07411
C5HC2 zinc finger / PHD-finger / ARID/BRIGHT DNA binding
C5HC2 zinc finger / PHD-finger / ARID/BRIGHT DNA binding
RUM1RUM1
150
150
04836
04836
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC10FZC10
151
151
00841
00841
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC40FZC40
152
152
06223
06223
MIZ/SP-RING zinc finger
MIZ/SP-RING zinc finger
MIZ1MIZ1
153
153
00830
00830
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC38FZC38
154
154
01551
01551
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT201GAT201
155
155
04908
04908
bZIP transcription factor / Basic region leucine zipper
bZIP transcription factor / Basic region leucine zipper
CLR4CLR4
상동 재조합 기법을 사용하여 178가지 중에서 155가지 잠재적인 전사인자 유전자를 결실시켰고, 이에 의해 전사인자의 기능을 분석하였다. 일련의 서명 표지를 포함하는 우성 노르세트로신 내성 표지자(nourseothricin-resistance markers, NATs)를 사용하여 대규모의 생체 내 독성 시험을 실시하였다.Homologous recombination techniques were used to delete 155 potential transcription factor genes out of 178, thereby analyzing the function of transcription factors. Large-scale in vivo toxicity tests were conducted using dominant norsetrosine-resistance markers (NATs) containing a series of signature markers.
서던 블롯에 의해 모든 전사인자 변이체 균주의 유전형을 검증하였고, 동시에 유전자 결실 및 전사인자 변이체 균주 내 유전자-파쇄 카세트의 전위성 병합이 없음을 확인하였다. 표현형을 정확하게 검증하고 관련되지 않은 변이 효과를 배제시키기 위해, 본 발명자는 기존에 알려진 4가지 전사인자(HXL1 , ATF1 , MBS1 및 SKN7(비특허문헌 9, 비특허문헌 10 및 비특허문헌 11)를 포함하는 155가지 전사인자에 대하여 두 가지 이상의 독립적인 전사인자 변이체(TF mutants)를 제작하였고 322가지 균주를 획득하였다(표 2 참조). Genotypes of all transcription factor variant strains were verified by Southern blot, and at the same time, it was confirmed that there was no gene deletion and translocation of the gene-breaking cassette in the transcription factor variant strain. In order to accurately verify the phenotype and exclude unrelated mutation effects, the present inventors use four previously known transcription factors ( HXL1 , ATF1 , MBS1, and SKN7 (
1
One
02566
02566
FKH2FKH2
FKH
FKH
YSB1339
YSB1339
2
2
YSB1340
YSB1340
3
3
00791
00791
HLH1HLH1
HLH
HLH
YSB1175
YSB1175
4
4
YSB1176
YSB1176
5
5
07464
07464
MBS1MBS1
APS
APS
YSB488
YSB488
6
6
YSB489
YSB489
7
7
03401
03401
GAT203GAT203
GAT
GAT
YSB569
YSB569
8
8
YSB570
YSB570
9
9
04588
04588
ERT1ERT1
FZC
FZC
YSB693
YSB693
10
10
YSB694
YSB694
11
11
00828
00828
SIP401SIP401
FZC
FZC
YSB1358
YSB1358
12
12
YSB1359
YSB1359
13
13
03561
03561
FZC33FZC33
FZC
FZC
YSB1074
YSB1074
14
14
YSB1075
YSB1075
15
15
06762
06762
GAT204GAT204
GAT
GAT
YSB1311
YSB1311
16
16
YSB1312
YSB1312
17
17
06276
06276
CEP3CEP3
FZC
FZC
YSB847
YSB847
18
18
YSB848
YSB848
19
19
05785
05785
STB4STB4
FZC
FZC
YSB1013
YSB1013
20
20
YSB1014
YSB1014
21
21
01438
01438
MBS2MBS2
APS
APS
YSB538
YSB538
22
22
YSB539
YSB539
23
23
07593
07593
YAP4YAP4
BZP
BZP
YSB1587
YSB1587
24
24
YSB1661
YSB1661
25
25
04837
04837
MLN1MLN1
HLH
HLH
YSB1172
YSB1172
26
26
YSB1173
YSB1173
27
27
05093
05093
HOB6HOB6
HOM
HOM
YSB1255
YSB1255
28
28
YSB1256
YSB1256
29
29
05642
05642
FZC37FZC37
FZC
FZC
YSB1329
YSB1329
30
30
YSB1330
YSB1330
31
31
05431
05431
RIM101RIM101
C2Z
C2Z
YSB1366
YSB1366
32
32
YSB1367
YSB1367
33
33
04398
04398
ARO80ARO80
FZC
FZC
YSB714
YSB714
34
34
YSB715
YSB715
35
35
04878
04878
FZC1FZC1
FZC
FZC
YSB510
YSB510
36
36
YSB511
YSB511
37
37
03183
03183
FZC24FZC24
FZC
FZC
YSB774
YSB774
38
38
YSB775
YSB775
39
39
03710
03710
ECM22ECM22
FZC
FZC
YSB476
YSB476
40
40
YSB478
YSB478
41
41
03279
03279
CCD4CCD4
HOM
HOM
YSB706
YSB706
42
42
YSB707
YSB707
43
43
04637
04637
MBF1MBF1
HTH
HTH
YSB768
YSB768
44
44
YSB769
YSB769
45
45
06425
06425
PPR1PPR1
FZC
FZC
YSB1046
YSB1046
46
46
YSB1047
YSB1047
47
47
04345
04345
ARO8001ARO8001
FZC
FZC
YSB661
YSB661
48
48
YSB662
YSB662
49
49
04184
04184
FZC47FZC47
FZC
FZC
YSB1406
YSB1406
50
50
YSB1407
YSB1407
51
51
02774
02774
MAL13MAL13
FZC
FZC
YSB506
YSB506
52
52
YSB507
YSB507
53
53
00670
00670
FZC12FZC12
FZC
FZC
YSB467
YSB467
54
54
YSB468
YSB468
55
55
05010
05010
ZFC7ZFC7
C2Z
C2Z
YSB481
YSB481
56
56
YSB482
YSB482
57
57
04090
04090
ATF1ATF1
BZP
BZP
YSB676
YSB676
58
58
YSB678
YSB678
59
59
06134
06134
BZP1BZP1
((
HXL1HXL1
))
BZP
BZP
YSB723
YSB723
60
60
YSB724
YSB724
61
61
04630
04630
YAP2YAP2
BZP
BZP
YSB1416
YSB1416
62
62
YSB1417
YSB1417
63
63
07901
07901
FZC29FZC29
FZC
FZC
YSB718
YSB718
64
64
YSB719
YSB719
65
65
03115
03115
FZC46FZC46
FZC
FZC
YSB1209
YSB1209
66
66
YSB1210
YSB1210
67
67
07924
07924
MCM1MCM1
SRF
SRF
YSB1302
YSB1302
68
68
YSB1303
YSB1303
69
69
07435
07435
HAP2HAP2
CCA
CCA
YSB1104
YSB1104
70
70
YSB1105
YSB1105
71
71
02555
02555
SIP402SIP402
FZC
FZC
YSB529
YSB529
72
72
YSB530
YSB530
73
73
04594
04594
FZC27FZC27
FZC
FZC
YSB582
YSB582
74
74
YSB583
YSB583
75
75
06188
06188
FZC15FZC15
FZC
FZC
YSB646
YSB646
76
76
YSB647
YSB647
77
77
05170
05170
PIP2PIP2
FZC
FZC
YSB1249
YSB1249
78
78
YSB1250
YSB1250
79
79
02241
02241
HOB5HOB5
HOM
HOM
YSB1585
YSB1585
80
80
YSB1586
YSB1586
81
81
06921
06921
HOB4HOB4
HOM
HOM
YSB1435
YSB1435
82
82
YSB1437
YSB1437
83
83
05186
05186
GRF1GRF1
GRF
GRF
YSB796
YSB796
84
84
YSB797
YSB797
85
85
00039
00039
ZFC6ZFC6
C2Z
C2Z
YSB1953
YSB1953
86
86
YSB1954
YSB1954
87
87
01454
01454
STE12STE12
C2Z
C2Z
YSB1542
YSB1542
88
88
YSB1543
YSB1543
89
89
03527
03527
HEL2HEL2
C2Z
C2Z
YSB1382
YSB1382
90
90
YSB1383
YSB1383
91
91
05255
05255
FZC2FZC2
FZC
FZC
YSB1050
YSB1050
92
92
YSB1051
YSB1051
93
93
01883
01883
GAT8GAT8
GAT
GAT
YSB471
YSB471
94
94
YSB472
YSB472
95
95
04353
04353
CLR1CLR1
C2Z
C2Z
YSB1396
YSB1396
96
96
YSB1397
YSB1397
97
97
03998
03998
RLM1RLM1
SRF
SRF
YSB1300
YSB1300
98
98
YSB1301
YSB1301
99
99
00239
00239
YAP1YAP1
BZP
BZP
YSB815
YSB815
100
100
YSB1290
YSB1290
101
101
06871
06871
FZC41FZC41
FZC
FZC
YSB1334
YSB1334
102
102
YSB1335
YSB1335
103
103
00156
00156
SP1(SP1(
CRZ1CRZ1
))
C2Z
C2Z
YSB1263
YSB1263
104
104
YSB1264
YSB1264
105
105
04268
04268
APN2APN2
GRF
GRF
YSB1429
YSB1429
106
106
YSB1430
YSB1430
107
107
05420
05420
USV101USV101
C2Z
C2Z
YSB1464
YSB1464
108
108
YSB1465
YSB1465
109
109
00018
00018
FZC6FZC6
FZC
FZC
YSB1980
YSB1980
110
110
YSB1981
YSB1981
111
111
03346
03346
BZP4BZP4
BZP
BZP
YSB1894
YSB1894
112
112
YSB1895
YSB1895
113
113
05538
05538
JJJ1JJJ1
C2Z
C2Z
YSB1592
YSB1592
114
114
YSB1594
YSB1594
115
115
03409
03409
SKN7SKN7
HSF
HSF
YSB349
YSB349
116
116
YSB350
YSB350
117
117
06339
06339
FZC35FZC35
FZC
FZC
YSB1341
YSB1341
118
118
YSB1342
YSB1342
119
119
07506
07506
FAP1FAP1
NFX
NFX
YSB813
YSB813
120
120
YSB817
YSB817
121
121
04807
04807
FZC8FZC8
FZC
FZC
YSB2112
YSB2112
122
122
YSB2113
YSB2113
123
123
02435
02435
BWC2BWC2
GAT
GAT
YSB1839
YSB1839
124
124
YSB1840
YSB1840
125
125
02364
02364
FZC19FZC19
FZC
FZC
YSB2115
YSB2115
126
126
YSB2116
YSB2116
127
127
03116
03116
HCM1HCM1
FKH
FKH
YSB1850
YSB1850
128
128
YSB1851
YSB1851
129
129
02877
02877
FZC51FZC51
FZC
FZC
YSB1842
YSB1842
130
130
YSB1843
YSB1843
131
131
00559
00559
BZP3BZP3
BZP
BZP
YSB1099
YSB1099
132
132
YSB1100
YSB1100
133
133
03914
03914
FZC14FZC14
FZC
FZC
YSB1846
YSB1846
134
134
YSB1847
YSB1847
135
135
00871
00871
CLR3CLR3
BZP
BZP
YSB1834
YSB1834
136
136
YSB1836
YSB1836
137
137
06483
06483
FZC25FZC25
FZC
FZC
YSB518
YSB518
138
138
YSB1822
YSB1822
139
139
07797
07797
CRL6CRL6
FKH
FKH
YSB1106
YSB1106
140
140
YSB1107
YSB1107
141
141
05019
05019
FZC21FZC21
FZC
FZC
YSB1252
YSB1252
142
142
YSB1253
YSB1253
143
143
05380
05380
FZC44FZC44
FZC
FZC
YSB2182
YSB2182
144
144
YSB2183
YSB2183
145
145
04518
04518
ZFC5ZFC5
C2Z
C2Z
YSB2177
YSB2177
146
146
YSB2178
YSB2178
147
147
05176
05176
HOB3HOB3
HOM
HOM
YSB2001
YSB2001
148
148
YSB2002
YSB2002
149
149
05861
05861
FKH101FKH101
FKH
FKH
YSB1855
YSB1855
150
150
YSB1856
YSB1856
151
151
00460
00460
LIV1LIV1
HLH
HLH
YSB2211
YSB2211
152
152
YSB2212
YSB2212
153
153
02305
02305
FZC45FZC45
FZC
FZC
YSB2221
YSB2221
154
154
YSB2222
YSB2222
155
155
03849
03849
ASG1ASG1
FZC
FZC
YSB3013
YSB3013
156
156
YSB3014
YSB3014
157
157
01014
01014
ZFC4ZFC4
C2Z
C2Z
YSB2231
YSB2231
158
158
YSB2232
YSB2232
159
159
01858
01858
HOB2HOB2
HOM
HOM
YSB2282
YSB2282
160
160
YSB2283
YSB2283
161
161
06719
06719
FZC49FZC49
FZC
FZC
YSB2171
YSB2171
162
162
YSB2173
YSB2173
163
163
04093
04093
YRM103YRM103
FZC
FZC
YSB2298
YSB2298
164
164
YSB2299
YSB2299
165
165
04176
04176
HSF2HSF2
HSF
HSF
YSB2295
YSB2295
166
166
YSB2296
YSB2296
167
167
01431
01431
HOB1HOB1
HOM
HOM
YSB2308
YSB2308
168
168
YSB2309
YSB2309
169
169
07443
07443
HLH4HLH4
HOM
HOM
YSB2244
YSB2244
170
170
YSB2245
YSB2245
171
171
04916
04916
FZC16FZC16
FZC
FZC
YSB2326
YSB2326
172
172
YSB2327
YSB2327
173
173
05392
05392
ZAP104ZAP104
C2Z
C2Z
YSB2134
YSB2134
174
174
YSB2135
YSB2135
175
175
02322
02322
FZC17FZC17
FZC
FZC
YSB2250
YSB2250
176
176
YSB2251
YSB2251
177
177
04012
04012
FZC18FZC18
FZC
FZC
YSB2320
YSB2320
178
178
YSB2321
YSB2321
179
179
00505
00505
FZC28FZC28
FZC
FZC
YSB2337
YSB2337
180
180
YSB2338
YSB2338
181
181
06751
06751
HLH3HLH3
HLH
HLH
YSB2329
YSB2329
182
182
YSB2330
YSB2330
183
183
04841
04841
FZC43FZC43
FZC
FZC
YSB517
YSB517
184
184
YSB2334
YSB2334
185
185
03212
03212
HCM101HCM101
FKH
FKH
YSB2390
YSB2390
186
186
YSB2391
YSB2391
187
187
01626
01626
ADA2ADA2
MYB
MYB
YSB2381
YSB2381
188
188
YSB2382
YSB2382
189
189
03894
03894
PDR802PDR802
FZC
FZC
YSB2387
YSB2387
190
190
YSB2388
YSB2388
191
191
02066
02066
FZC13FZC13
FZC
FZC
YSB2517
YSB2517
192
192
YSB2518
YSB2518
193
193
06818
06818
HAP1HAP1
FZC
FZC
YSB2481
YSB2481
194
194
YSB2482
YSB2482
195
195
05940
05940
ZFC3ZFC3
C2Z
C2Z
YSB2108
YSB2108
196
196
YSB2386
YSB2386
197
197
01948
01948
FZC36FZC36
FZC
FZC
YSB2335
YSB2335
198
198
YSB2523
YSB2523
199
199
04804
04804
SRE1SRE1
HLH
HLH
YSB2493
YSB2493
200
200
YSB2494
YSB2494
201
201
04352
04352
ZAP103ZAP103
C2Z
C2Z
YSB2540
YSB2540
202
202
YSB2541
YSB2541
203
203
03768
03768
FZC32FZC32
FZC
FZC
YSB2385
YSB2385
204
204
YSB2526
YSB2526
205
205
04895
04895
FZC3FZC3
FZC
FZC
YSB2611
YSB2611
206
206
YSB2664
YSB2664
207
207
04583
04583
DDT1DDT1
DDT
DDT
YSB1583
YSB1583
208
208
YSB2633
YSB2633
209
209
01973
01973
ZFC2ZFC2
C2Z
C2Z
YSB2622
YSB2622
210
210
YSB2623
YSB2623
211
211
06156
06156
FZC7FZC7
FZC
FZC
YSB2704
YSB2704
212
212
YSB2705
YSB2705
213
213
03431
03431
FZC48FZC48
FZC
FZC
YSB2646
YSB2646
214
214
YSB2647
YSB2647
215
215
07922
07922
FZC4FZC4
FZC
FZC
YSB2724
YSB2724
216
216
YSB2725
YSB2725
217
217
00031
00031
MLR1MLR1
FZC
FZC
YSB2727
YSB2727
218
218
YSB2728
YSB2728
219
219
03018
03018
ASG101ASG101
FZC
FZC
YSB2697
YSB2697
220
220
YSB2698
YSB2698
221
221
04263
04263
BZP2BZP2
BZP
BZP
YSB2702
YSB2702
222
222
YSB2703
YSB2703
223
223
01708
01708
GAT7GAT7
GAT
GAT
YSB2699
YSB2699
224
224
YSB2700
YSB2700
225
225
00332
00332
SIP4SIP4
FZC
FZC
YSB2680
YSB2680
226
226
YSB2681
YSB2681
227
227
07724
07724
CUF1CUF1
CDB
CDB
YSB2665
YSB2665
228
228
YSB2666
YSB2666
229
229
03902
03902
RDS2RDS2
FZC
FZC
YSB1898
YSB1898
230
230
YSB1899
YSB1899
231
231
03366
03366
ZNF2ZNF2
C2Z
C2Z
YSB2740
YSB2740
232
232
YSB2741
YSB2741
233
233
05222
05222
NRG1NRG1
C2Z
C2Z
YSB3096
YSB3096
234
234
YSB3097
YSB3097
235
235
05049
05049
PIP201PIP201
FZC
FZC
YSB3099
YSB3099
236
236
YSB3100
YSB3100
237
237
02516
02516
HLH5HLH5
HLH
HLH
YSB2609
YSB2609
238
238
YSB3059
YSB3059
239
239
04774
04774
FZC26FZC26
FZC
FZC
YSB3084
YSB3084
240
240
YSB3085
YSB3085
241
241
03336
03336
FZC50FZC50
FZC
FZC
YSB3131
YSB3131
242
242
YSB3132
YSB3132
243
243
03086
03086
FZC20FZC20
FZC
FZC
YSB3128
YSB3128
244
244
YSB3129
YSB3129
245
245
03229
03229
YOX101YOX101
HOM
HOM
YSB3134
YSB3134
246
246
YSB3136
YSB3136
247
247
01841
01841
GLN3GLN3
GAT
GAT
YSB3154
YSB3154
248
248
YSB3155
YSB3155
249
249
02476
02476
YRM101YRM101
FZC
FZC
YSB2997
YSB2997
250
250
YSB2998
YSB2998
251
251
05153
05153
GAT5GAT5
GAT
GAT
YSB3033
YSB3033
252
252
YSB3034
YSB3034
253
253
02700
02700
ZFC8ZFC8
C2Z
C2Z
YSB3031
YSB3031
254
254
YSB3032
YSB3032
255
255
04586
04586
HOB7HOB7
HOM
HOM
YSB3026
YSB3026
256
256
YSB3027
YSB3027
257
257
02723
02723
FZC23FZC23
FZC
FZC
YSB3105
YSB3105
258
258
YSB3106
YSB3106
259
259
03741
03741
FZC31FZC31
FZC
FZC
YSB3093
YSB3093
260
260
YSB3094
YSB3094
261
261
04457
04457
FZC30FZC30
FZC
FZC
YSB2447
YSB2447
262
262
YSB2448
YSB2448
263
263
04836
04836
FZC10FZC10
FZC
FZC
YSB3083
YSB3083
264
264
YSB3368
YSB3368
265
265
06223
06223
MIZ1MIZ1
MIZ
MIZ
YSB2133
YSB2133
266
266
YSB3366
YSB3366
267
267
01551
01551
GAT201GAT201
GAT
GAT
YSB3300
YSB3300
268
268
YSB3301
YSB3301
269
269
04908
04908
CLR4CLR4
BZP
BZP
YSB3282
YSB3282
270
270
YSB3283
YSB3283
271
271
07940
07940
BZP5BZP5
BZP
BZP
YSB1474
YSB1474
272
272
YSB1475
YSB1475
273
273
05112
05112
FZC42FZC42
FZC
FZC
YSB687
YSB687
274
274
YSB690
YSB690
275
275
00193
00193
GAT1GAT1
GAT
GAT
YSB2972
YSB2972
276
276
YSB2973
YSB2973
277
277
06814
06814
SXI1alphaSXI1alpha
HOM
HOM
YSB1390
YSB1390
278
278
YSB1391
YSB1391
279
279
YSB1392
YSB1392
280
280
00896
00896
FZC34FZC34
FZC
FZC
YSB501
YSB501
281
281
YSB2979
YSB2979
282
282
07411
07411
RUM1RUM1
PHZ
PHZ
YSB3164
YSB3164
283
283
YSB3747
YSB3747
284
284
00830
00830
FZC38FZC38
FZC
FZC
YSB777
YSB777
285
285
YSB3791
YSB3791
286
286
01069
01069
FZC11FZC11
FZC
FZC
YSB845
YSB845
287
287
YSB846
YSB846
288
288
YSB2983
YSB2983
289
289
03132
03132
FZC5FZC5
FZC
FZC
YSB1400
YSB1400
290
290
YSB1401
YSB1401
291
291
YSB1404
YSB1404
292
292
00068
00068
MET32MET32
C2Z
C2Z
YSB1178
YSB1178
293
293
YSB1179
YSB1179
294
294
YSB1180
YSB1180
295
295
01173
01173
PAN1PAN1
P53
P53
YSB1181
YSB1181
296
296
YSB1182
YSB1182
297
297
YSB1183
YSB1183
298
298
00514
00514
GAT6GAT6
GAT
GAT
YSB1384
YSB1384
299
299
YSB1385
YSB1385
300
300
YSB1386
YSB1386
301
301
07011
07011
FZC22FZC22
FZC
FZC
YSB1688
YSB1688
302
302
YSB1689
YSB1689
303
303
YSB2974
YSB2974
304
304
02603
02603
ZFC1ZFC1
C2Z
C2Z
YSB2573
YSB2573
305
305
YSB2574
YSB2574
306
306
YSB2575
YSB2575
307
307
04036
04036
HSF3HSF3
HSF
HSF
YSB2527
YSB2527
308
308
YSB2528
YSB2528
309
309
YSB2529
YSB2529
310
310
03059
03059
FZC9FZC9
FZC
FZC
YSB2984
YSB2984
311
311
YSB3266
YSB3266
312
312
YSB3267
YSB3267
313
313
06283
06283
LIV4LIV4
MYB
MYB
YSB2089
YSB2089
314
314
YSB3755
YSB3755
315
315
YSB3756
YSB3756
316
316
00841
00841
FZC40FZC40
FZC
FZC
YSB3088
YSB3088
317
317
YSB3758
YSB3758
318
318
01977
01977
FZC39FZC39
FZC
FZC
YSB1820
YSB1820
319
319
YSB2621
YSB2621
320
320
05375
05375
HLH2HLH2
HLH
HLH
YSB1147
YSB1147
321
321
YSB1148
YSB1148
322
322
YSB1149
YSB1149
CMO18 균주(독성이 약한 H99 균주, 비특허문헌 12)에서, 기존에 결실되어 2가지 이상의 독립적인 변이체로부터 유래된 53가지 전사인자를 결실시켜서 병렬적인 생체 외(in vitro) 및 생체 내(in vivo) 표현형 분석을 실시하였다. RIM101, ADA2, CUF1, SXL1, SP -1/ CRZ1, NRG1, STE12, BWC2, SRE1, ZNF2 및 HAP1/HAP2를 포함하는 알려진 전사인자들은 독립적으로 결실시켜서 표현형을 정확하게 비교하였다. 2가지 독립적인 전사인자 변이체 균주가 일치하지 않는 표현형을 나타내는 경우, 추가적인 전사인자 변이체를 제작하였고 기존의 변이체는 배제하였다. 본 발명에서 유전자 녹아웃 중 약 8%(13가지 전사인자)는 일치하지 않는 표현형을 나타내는 것으로 확인되었는데, 유전체 내 예상치 못한 변이에 의한 것으로 예측되었다. 이러한 수준(7%)은 자낭균 진균 병원체 캔디다 알비칸스(비특허문헌 13)에 대한 선행연구 결과와 매우 유사하였다. 나머지 23가지 전사인자에 대해서는 전사인자 변이체를 제작하지 않았다. 최종적으로, 크립토코커스 네오포만스 균주 내 155가지 전사인자에 대하여 322가지 전사인자 변이체 균주를 제작하였다(도 3b).In the CMO18 strain (low toxicity H99 strain, Non-Patent Document 12), 53 transcription factors derived from two or more independent mutants were previously deleted, and thus parallel in vitro and in vivo ) Phenotypic analysis was performed. RIM101 , ADA2 , CUF1 , SXL1 , SP -1/ CRZ1 , NRG1 , STE12 , BWC2 , SRE1 , Known transcription factors including ZNF2 and HAP1 / HAP2 were independently deleted to accurately compare phenotypes. When two independent transcription factor variant strains exhibit inconsistent phenotypes, additional transcription factor variants were constructed and existing variants were excluded. In the present invention, about 8% of gene knockouts (13 transcription factors) were found to exhibit inconsistent phenotypes, which were predicted to be due to unexpected mutations in the genome. This level (7%) was very similar to the results of previous studies on Candida albicans (Non-Patent Literature 13), a pathogen of ascytic fungi. For the remaining 23 transcription factors, transcription factor variants were not produced. Finally, 322 transcription factor variant strains were prepared for 155 transcription factors in the Cryptococcus neoformans strain (FIG. 3B).
전사인사 변이체에 대한 156가지 전사인자 중에서, 58가지 유전자는 FungiDB(www.fungidb.org) 내 등록되거나 기존에 공개된 연구에 사용된 명칭을 사용하였다. 나머지 98가지 전사인자에 대하여, 최근에 개시된 크립토코커스 네오포만스 균주 내 명명법에 따라서 유전자 이름을 명명하였다(비특허문헌 14).Among the 156 transcription factors for the transcriptional personnel variant, 58 genes were registered in FungiDB (www.fungidb.org) or used in previously published studies. For the remaining 98 transcription factors, the gene names were named according to the nomenclature within the recently disclosed Cryptococcus neoformans strain (Non-Patent Document 14).
1.2 전사인자 1.2 transcription factor 변이체Variant 제작 making
크립토코커스 네오포만스(C. neoformans) 혈청형 A H99S 균주를 배경으로 크립토코커스 네오포만스 균주의 전사인자 결실변이체(이하, TFKO)를 제작하였다. 노르세트로신 내성 표지자(nourseothricin-resistance marker, NAT) 및 서명-태그 서열를 포함하는 유전자 파쇄 카세트는 서열목록 및 표 2에 기재된 프라이머 세트를 이용한 중복(overlap) 중합효소연쇄반응(이하, PCR) 또는 더블-조인트(double-joint) PCR에 의해 제작하였다(비특허문헌 15 및 비특허문헌 16). 중복 PCR과정 중에, 전사인자 유전자의 5' 및 3' 인접 영역은 PCR 1차 라운드(round)에서 H99 게놈 DNA에 대하여 각각 프라이머 L1 및 R1, 프라이머 L2 및 R2(표 1 참조)를 사용하여 증폭하였다. 프라이머 M13Fe(M13 정방향 확장) 및 프라이머 M13Re(M13 역방향 확장)를 사용하여 독특한 내성 표지자 서열을 포함하는 우성 선별 마커(NATr)를 증폭하였다. PCR 2차 라운드에서, 전사인자 유전자-파쇄 카세트는 1-라운드 PCR 생성물을 주형으로 하여 프라이머 L1 및 R2를 사용하여 중복 PCR을 수행하여 제작하였다. 더블 조인트 PCR 방법에서, 전사인자 유전자의 5' 및 3' 인접 영역은 PCR 1라운드 내 H99 게놈 DNA와 프라이머 쌍 L1/L2 및 R1/R2를 각각 사용하여 증폭하였다. NAT 스플리트 마커의 5' 및 3' 영역은 독특한 서명-태그 서열을 포함하는 pNATSTM(듀크대학교 Joeseph Heitman 연구실로부터 입수)에 대하여 프라이머 M13Fe, NSL, 프라이머 M13Re, 및 NSR을 사용하여 증폭하였다. 증폭시킨 유전자-파쇄 카세트는 600 ug의 미세운반 금 비드(0.6 um, Bio-Rad)와 결합시켰고, 2.5M 염화칼슘 10 ㎕ 및 1 M 스퍼미딘 2 ㎕을 처리하였다. 유전자-파쇄 카세트를 결합시킨 금 비드는 바이오리스틱(biolistic) 형질전환 장치(비특허문헌 17)을 사용하여 H99S 균주(듀크대학교 Joeseph Heitman 연구실로부터 입수)에 도입하였다. 회복에 필요한 4시간 배양 후에, 세포를 긁어내어 노르세트로신(100 ㎍/㎖)을 포함하는 효모 추출물-펩톤 덱스트로즈(이하, YPD) 배지 상에 옮기고 이후에 30℃에서 3일 내지 5일간 배양하였다. 안정적인 노르세트로신-내성 형질전환체는 표 3 및 서열목록에 기재된 프라이머 세트를 이용한 진단 PCR을 실시하여 스크리닝하였다.A transcription factor deletion variant (hereinafter, TFKO) of the Cryptococcus neoformans strain was prepared with the C. neoformans serotype A H99S strain in the background. The gene disruption cassette including the norseothricin-resistance marker (NAT) and the signature-tag sequence is an overlap polymerase chain reaction (hereinafter, PCR) or It was produced by double-joint PCR (
모든 전사인자 변이체는 한국미생물보존센터(KCCM) 및 미국 듀크대 미생물 발병 센터에 기탁하였다.All transcription factor variants were deposited with the Korea Microbial Conservation Center (KCCM) and the Microbial Outbreak Center at Duke University in the United States.
표현형 프로파일링(profiling)Phenotypic profiling
제작된 322가지 전사인자 변이체에 대하여, 다양한 표현형(성장, 분화, 형태학, 스트레스 반응, 항균제 내성, 독성-인자 생산 및 생체 내 독성)에 대한 생체 내(in vivo) 및 생체 외(in vitro) 표현형 분석을 실시하였다(도 1b). 모든 표현형 데이터 세트는 수색 표준으로 표현하였는데, 6가지 다른 성장 조건에서 RNA-seq 분석에 의해 측정된 각 전사인자의 전사체 수준을 측정하였다(도 2). 열 지도 내의 적색 및 청색은 각각 감소 및 증가를 나타내었다. 표현형 강도(강, 중, 약)는 적색 또는 청색의 증감률로 나타내었다. 표현형 분석 결과는 전사인자 변이체의 약 93%(145/155)가 최소한 하나의 특별한 표현형을 나타냄을 증명하였는데, 이는 전사인자 변이체 집단의 높은 기능성을 제시하였다. 표현형 분석 결과는 전사인자 변이체의 대략 93%(145/155)가 최소한 하나의 특징적인 표현형을 나타낸다는 것을 검증하였다. 전사인자의 85%(132/155)는 기존에는 크립토코커스 네오포만스 균주 내에서의 기능이 알려지지 않은 유전자이다. For the 322 transcription factor variants produced, in vivo and in vitro phenotypes for various phenotypes (growth, differentiation, morphology, stress response, antimicrobial resistance, toxicity-factor production and in vivo toxicity) Analysis was carried out (Fig. 1B). All phenotypic data sets were expressed as search standards, and the transcript levels of each transcription factor were measured by RNA-seq analysis under six different growth conditions (FIG. 2). Red and blue color in the heat map indicated decrease and increase, respectively. The phenotypic intensity (strong, medium, weak) was expressed as the rate of increase or decrease in red or blue. The results of phenotypic analysis proved that about 93% (145/155) of the transcription factor variants exhibited at least one specific phenotype, which suggested high functionality of the transcription factor variant population. Phenotypic analysis results verified that approximately 93% (145/155) of the transcription factor variants exhibited at least one characteristic phenotype. 85% (132/155) of transcription factors are genes whose function in the Cryptococcus neoformans strain was previously unknown.
발형체(phenome)에 대한 모든 자료는 크립토코커스 네오포만스 전사인자 데이터베이스 내에 공개되어 사용할 수 있다(http://tf.cryptococcus.org).All data on the phenome are publicly available and available in the Cryptococcus neoformans transcription factor database (http://tf.cryptococcus.org).
2.1 유전형 분석2.1 Genotyping
양성 형질변이체의 정확한 유전형은 서던 블롯(southern blot) 분석에 의해 검증하였다. 크립토코커스 게놈 DNA는 세틸 트리메틸 암모늄 브로마이드(CTAB,Cetyl Trimethyl Ammonium Bromide) 방법(비특허문헌 33)을 사용하여 추출하였다. 각각의 TFKO 변이체로부터 추출한 게놈 DNA는 지정된 제한 효소를 사용하여 절단하였다(표 4 참조). 절단된 게놈 DNA는 1% 아가로즈 젤-전기영동에 의해 분리하였다. 아가로즈 젤은 0.5 M 수산화나트륨 및 1.5 M 염화나트륨을 포함하는 변성 완충액으로 옮겨서 45분간 방치하였다. 이후에, 아가로즈 젤은 1.5 M 염화나트륨 및 pH 8로 적정된 0.5 M 트리스 완충액을 포함하는 중성 완충액으로 옮겨서 45분간 방치하였다. 절단된 게놈 DNA는 10배 SSC(Saline-Sodium Citrate) 완충액을 이용하여 나일론 멤브레인으로 옮겼고 1200 J/m2의 자외선에 노출하여 고정시켰다. 멤브레인은 변형된 처치(church) 혼성화 완충액(1 mM EDTA, 0.25 M Na2HPO4, 1 %의 가수분해된 카제인, 7% SDS, 6% H3PO4)을 사용하여 유전자 특이적이면서 방사성 활성 표지된 탐침과 혼성화시켰다. 멤브레인은 세척 완충액 1(2X SSC 및 0.1% SDS 포함) 및 세척 완충액2 (1X SSC 및 0.1% SDS 포함)로 15분간 세척하였다. 이후에, 멤브레인은 오토그래피(autography) 필름에 1일간 노출시켰다.The correct genotype of the positive mutant was verified by Southern blot analysis. Cryptococcus genomic DNA was extracted using the Cetyl Trimethyl Ammonium Bromide (CTAB) method (Non-Patent Document 33). Genomic DNA extracted from each TFKO variant was digested using a designated restriction enzyme (see Table 4). The cut genomic DNA was separated by 1% agarose gel-electrophoresis. The agarose gel was transferred to a denaturation buffer containing 0.5 M sodium hydroxide and 1.5 M sodium chloride and left for 45 minutes. Thereafter, the agarose gel was transferred to a neutral buffer containing 1.5 M sodium chloride and 0.5 M Tris buffer titrated to
division
H99 lD
H99 lD
Designated name
Designated name
Restriction enzyme cut
Restriction enzyme cut
1
One
02566
02566
FKH2FKH2
BamHI
BamHI
2
2
00791
00791
HLH1HLH1
BglII
BglII
3
3
01069
01069
FZC11FZC11
SphI
SphI
4
4
07464
07464
MBS1MBS1
SphI
SphI
5
5
03401
03401
GAT203GAT203
HindIII
HindIII
6
6
04588
04588
ERT1ERT1
EcoRV
EcoRV
7
7
00828
00828
SIP401SIP401
EcoRI
EcoRI
8
8
03561
03561
FZC33FZC33
EcoRV
EcoRV
9
9
06762
06762
GAT204GAT204
KpnI
KpnI
10
10
06276
06276
CEP3CEP3
SphI
SphI
11
11
05785
05785
STB4STB4
BamHI
BamHI
12
12
03132
03132
FZC5FZC5
PstI
PstI
13
13
01438
01438
MBS2MBS2
EcoRI
EcoRI
14
14
07593
07593
YAP4YAP4
SmaI
SmaI
15
15
04837
04837
MLN1MLN1
SmaI
SmaI
16
16
05093
05093
HOB6HOB6
KpnI
KpnI
17
17
05642
05642
FZC37FZC37
XmaI
XmaI
18
18
05431
05431
RIM101RIM101
ClaI
ClaI
19
19
04398
04398
ARO80ARO80
HincII
HincII
20
20
04878
04878
FZC1FZC1
EcoRI
EcoRI
21
21
03183
03183
FZC24FZC24
EcoRV
EcoRV
22
22
03710
03710
ECM22ECM22
HindIII
HindIII
23
23
03279
03279
CCD4CCD4
HindIII
HindIII
24
24
04637
04637
MBF1MBF1
HindIII
HindIII
25
25
06425
06425
PPR1PPR1
EcoRV
EcoRV
26
26
04345
04345
ARO8001ARO8001
BamHI
BamHI
27
27
04184
04184
FZC47FZC47
BglII
BglII
28
28
02774
02774
MAL13MAL13
EcoRV
EcoRV
29
29
00670
00670
FZC12FZC12
StyI
StyI
30
30
00068
00068
MET32MET32
AfeI
AfeI
31
31
05010
05010
ZFC7ZFC7
ScaI
ScaI
32
32
04090
04090
ATF1ATF1
PstI
PstI
33
33
06134
06134
BZP1BZP1
((
HXL1HXL1
))
BamHI
BamHI
34
34
04630
04630
YAP2YAP2
HindIII
HindIII
35
35
07901
07901
FZC29FZC29
EcoRV
EcoRV
36
36
01173
01173
PAN1PAN1
SalI
SalI
37
37
03115
03115
FZC46FZC46
SacI
SacI
38
38
07924
07924
MCM1MCM1
XbaI
XbaI
39
39
07435
07435
HAP2HAP2
SalI
SalI
40
40
02555
02555
SIP402SIP402
MfeI
MfeI
41
41
04594
04594
FZC27FZC27
HindIII
HindIII
42
42
06188
06188
FZC15FZC15
BamHI
BamHI
43
43
05170
05170
PIP2PIP2
BamHI
BamHI
44
44
02241
02241
HOB5HOB5
PstI
PstI
45
45
06921
06921
HOB4HOB4
KpnI
KpnI
46
46
05186
05186
GRF1GRF1
EcoRI
EcoRI
47
47
00896
00896
FZC34FZC34
HindIII
HindIII
48
48
00039
00039
ZFC6ZFC6
KpnI
KpnI
49
49
07940
07940
BZP5BZP5
BamHI
BamHI
50
50
06814
06814
SXI1alphaSXI1alpha
SacI
SacI
51
51
01454
01454
STE12STE12
BamHI
BamHI
52
52
03527
03527
HEL2HEL2
BamHI
BamHI
53
53
05255
05255
FZC2FZC2
HindIII
HindIII
54
54
05112
05112
FZC42FZC42
BglII, EcoRV
BglII, EcoRV
55
55
01883
01883
GAT8GAT8
SacI
SacI
56
56
04353
04353
CLR1CLR1
SalI
SalI
57
57
05375
05375
HLH2HLH2
HindIII
HindIII
58
58
03998
03998
RLM1RLM1
StyI
StyI
59
59
00239
00239
YAP1YAP1
BamHI
BamHI
60
60
06871
06871
FZC41FZC41
BamHI, XbaI
BamHI, XbaI
61
61
00156
00156
SP1(SP1(
CRZ1CRZ1
))
EcoRV
EcoRV
62
62
04268
04268
APN2APN2
EcoRV
EcoRV
63
63
05420
05420
USV101USV101
SphI
SphI
64
64
00018
00018
FZC6FZC6
SphI
SphI
65
65
03346
03346
BZP4BZP4
PstI
PstI
66
66
00514
00514
GAT6GAT6
EcoRV
EcoRV
67
67
05538
05538
JJJ1JJJ1
SphI
SphI
68
68
03409
03409
SKN7SKN7
BamHI
BamHI
69
69
06339
06339
FZC35FZC35
HindIII
HindIII
70
70
07011
07011
FZC22FZC22
PstI
PstI
71
71
07506
07506
FAP1FAP1
HindIII
HindIII
72
72
04807
04807
FZC8FZC8
PstI
PstI
73
73
02435
02435
BWC2BWC2
HindIII
HindIII
74
74
02364
02364
FZC19FZC19
BamHI
BamHI
75
75
03116
03116
HCM1HCM1
SphI
SphI
76
76
02877
02877
FZC51FZC51
SphI
SphI
77
77
00559
00559
BZP3BZP3
SphI
SphI
78
78
03914
03914
FZC14FZC14
PstI
PstI
79
79
00871
00871
CLR3CLR3
XbaI
XbaI
80
80
06483
06483
FZC25FZC25
PstI
PstI
81
81
07797
07797
CRL6CRL6
EcoRI
EcoRI
82
82
05019
05019
FZC21FZC21
PstI
PstI
83
83
05380
05380
FZC44FZC44
SacI
SacI
84
84
04518
04518
ZFC5ZFC5
EcoRI
EcoRI
85
85
05176
05176
HOB3HOB3
XhoI
XhoI
86
86
05861
05861
FKH101FKH101
EcoRV
EcoRV
87
87
00460
00460
LIV1LIV1
SphI
SphI
88
88
02305
02305
FZC45FZC45
EcoRV
EcoRV
89
89
03849
03849
ASG1ASG1
KpnI
KpnI
90
90
01014
01014
ZFC4ZFC4
SalI
SalI
91
91
01858
01858
HOB2HOB2
SphI
SphI
92
92
06719
06719
FZC49FZC49
PstI
PstI
93
93
04093
04093
YRM103YRM103
SphI
SphI
94
94
04176
04176
HSF2HSF2
KpnI
KpnI
95
95
01431
01431
HOB1HOB1
HindIII
HindIII
96
96
07443
07443
HLH4HLH4
SphI
SphI
97
97
04916
04916
FZC16FZC16
EcoRI
EcoRI
98
98
05392
05392
ZAP104ZAP104
SalI
SalI
99
99
02322
02322
FZC17FZC17
XbaI
XbaI
100
100
04012
04012
FZC18FZC18
HindIII
HindIII
101
101
00505
00505
FZC28FZC28
SmaI
SmaI
102
102
06751
06751
HLH3HLH3
SalI
SalI
103
103
04841
04841
FZC43FZC43
XbaI
XbaI
104
104
03212
03212
HCM101HCM101
SacI, SalI
SacI, SalI
105
105
01626
01626
ADA2ADA2
BamHI
BamHI
106
106
03894
03894
PDR802PDR802
SphI
SphI
107
107
02066
02066
FZC13FZC13
BamHI
BamHI
108
108
06818
06818
HAP1HAP1
HindIII
HindIII
109
109
05940
05940
ZFC3ZFC3
EcoRI
EcoRI
110
110
01948
01948
FZC36FZC36
XhoI
XhoI
111
111
04804
04804
SRE1SRE1
BamHI
BamHI
112
112
04352
04352
ZAP103ZAP103
SacI, SalI
SacI, SalI
113
113
03768
03768
FZC32FZC32
BamHI
BamHI
114
114
01977
01977
FZC39FZC39
KpnI
KpnI
115
115
04895
04895
FZC3FZC3
KpnI
KpnI
116
116
04583
04583
DDT1DDT1
SphI
SphI
117
117
01973
01973
ZFC2ZFC2
SphI
SphI
118
118
02603
02603
ZFC1ZFC1
HindIII
HindIII
119
119
04036
04036
HSF3HSF3
SphI
SphI
120
120
06156
06156
FZC7FZC7
EcoRI
EcoRI
121
121
03431
03431
FZC48FZC48
EcoRV
EcoRV
122
122
07922
07922
FZC4FZC4
EcoRV
EcoRV
123
123
00031
00031
MLR1MLR1
PstI, XmaI
PstI, XmaI
124
124
03018
03018
ASG101ASG101
XhoI
XhoI
125
125
04263
04263
BZP2BZP2
EcoRV
EcoRV
126
126
01708
01708
GAT7GAT7
SacI
SacI
127
127
00332
00332
SIP4SIP4
PstI
PstI
128
128
07724
07724
CUF1CUF1
KpnI
KpnI
129
129
03902
03902
RDS2RDS2
EcoRV, EcoRI
EcoRV, EcoRI
130
130
03366
03366
ZNF2ZNF2
EcoRV
EcoRV
131
131
05222
05222
NRG1NRG1
EcoRV
EcoRV
132
132
05049
05049
PIP201PIP201
BamHI
BamHI
133
133
02516
02516
HLH5HLH5
BamHI
BamHI
134
134
04774
04774
FZC26FZC26
PstI
PstI
135
135
03059
03059
FZC9FZC9
EcoRV
EcoRV
136
136
00193
00193
GAT1GAT1
SphI
SphI
137
137
03336
03336
FZC50FZC50
HindIII
HindIII
138
138
03086
03086
FZC20FZC20
KpnI
KpnI
139
139
03229
03229
YOX101YOX101
XbaI
XbaI
140
140
01841
01841
GLN3GLN3
EcoRI
EcoRI
141
141
02476
02476
YRM101YRM101
XhoI
XhoI
142
142
05153
05153
GAT5GAT5
SalI
SalI
143
143
02700
02700
ZFC8ZFC8
EcoRV
EcoRV
144
144
04586
04586
HOB7HOB7
SphI
SphI
145
145
02723
02723
FZC23FZC23
EcoRV
EcoRV
146
146
03741
03741
FZC31FZC31
ClaI
ClaI
147
147
04457
04457
FZC30FZC30
BamHI
BamHI
148
148
06283
06283
LIV4LIV4
EcoRV
EcoRV
149
149
07411
07411
RUM1RUM1
EcoRV
EcoRV
150
150
04836
04836
FZC10FZC10
EcoRV
EcoRV
151
151
00841
00841
FZC40FZC40
EcoRI
EcoRI
152
152
06223
06223
MIZ1MIZ1
EcoRV
EcoRV
153
153
00830
00830
FZC38FZC38
SphI
SphI
154
154
01551
01551
GAT201GAT201
XbaI
XbaI
155
155
04908
04908
CLR4CLR4
HindIII
HindIII
2.2 유전자 발현 분석2.2 Gene expression analysis
유전자 발현은 노던 블롯(northern blot)을 사용하여 분석하였다. 전체 RNA는 트라이졸을 사용하여 각각의 시료로부터 추출하였다. RNA 10 ㎍은 DEPC(diethyl pryocarbonate)-처리한 물을 포함하는 1% 아가로스 젤 및 1 x MOPS(3-(N-morpholino) propane sulfonic acid) 러닝 완충액 내에서 전기영동에 의해 분리시켰다. 젤은 증류수로 3회 세척하였고, 20 x SSC 완충액을 사용하여 나일론 멤브레인(Millipore, INYC00010)으로 옮겼고 1200 J/m2 자외선에 노출하여 고정시켰다. 멤브레인은 변형된 처치(church) 혼성화 완충액(1mM EDTA (바이오세상, E1002), 0.25M Na2HPO4 (Sigma, S9763), 1% N-2-Amine (Sigma, C0626), 7% SDS (Bioshop, SDS001), 0.17% H3PO4 (Sigma, #438081))을 사용하여 유전자 특이적이며 방사선 활성 표지된 탐침과 혼성화하였다. 멤브레인은 세척 완충액 1(2X SSC 완충액, 0.1% SDS) 및 세척 완충액 2(1X SSC 완충액, 0.1% SDS)를 사용하여 세척하였고, 이후에 멤브레인은 오토그래피 필름에 1 내지 2일간 노출시켰다.Gene expression was analyzed using Northern blot. Total RNA was extracted from each sample using Trizol. 10 μg of RNA was separated by electrophoresis in 1% agarose gel containing DEPC (diethyl pryocarbonate)-treated water and 1 x MOPS (3-(N-morpholino) propane sulfonic acid) running buffer. The gel was washed 3 times with distilled water, transferred to a nylon membrane (Millipore, INYC00010) using 20 x SSC buffer, and fixed by exposure to 1200 J/m 2 ultraviolet rays. Membrane is modified church hybridization buffer (1mM EDTA ( Biosesang, E1002), 0.25M Na 2 HPO 4 (Sigma, S9763), 1% N-2-Amine (Sigma, C0626), 7% SDS (Bioshop , SDS001), 0.17% H 3 PO 4 (Sigma, #438081)) was used to hybridize with a gene-specific and radioactive labeled probe. The membrane was washed with wash buffer 1 (2X SSC buffer, 0.1% SDS) and wash buffer 2 (1X SSC buffer, 0.1% SDS), and then the membrane was exposed to the autograph film for 1-2 days.
성장을 조절하는 전사 인자(Transcription factors that regulate growth ( TFTF ))
크립토코커스 네오포만스 균주는 자연 상태 및 동물 숙주 환경 내에서 부생균 및 기생의 생활 주기를 갖고 있다. 따라서, 크립토코커스 네오포만스 균주는 상온(25℃) 내지 고온(37℃~39℃) 범위의 온도에서 성장할 수 있어야 한다. 다양한 온도에서의 전사인자 변이체 균주의 성장 표현형을 분석하기 위해서, 효모 추출물-펩톤 덱스트로즈(YPD) 배지[Yeast extract (Becton, Dickison and company #212750), Peptone (Becton, Dickison and company #211677), Glucose (Duchefa, #G0802)] 상의 다양한 온도 범위(25℃, 30℃, 37℃ 및 39℃)에서 각각 변이체의 성장을 관찰하였다. 일부 전사인자(BZP2, CUF1, LIV4, GAT5, FZC6, NRG1)의 결실은 온도에 무관한 성장-결핍을 나타내었다(도 3). 외부의 CuSO4 투여에 의해 cuf1 야생형 성장이 회복되므로, cuf1 변이체의 성장 결핍은 구리 흡수의 장애에 의한 것이다.Cryptococcus neoformans strains have a life cycle of by-products and parasitics in their natural state and in the animal host environment. Therefore, the Cryptococcus neoformans strain should be able to grow at a temperature ranging from room temperature (25°C) to high temperature (37°C to 39°C). In order to analyze the growth phenotype of the transcription factor variant strain at various temperatures, yeast extract-peptone dextrose (YPD) medium (Yeast extract (Becton, Dickison and company #212750), Peptone (Becton, Dickison and company #211677) , Glucose (Duchefa, #G0802)] in various temperature ranges (25 ℃, 30 ℃, 37 ℃ and 39 ℃) was observed the growth of the mutant, respectively. Deletion of some transcription factors (BZP2, CUF1, LIV4, GAT5, FZC6, NRG1) showed growth-deficiency independent of temperature (FIG. 3). External CuSO 4 Since the cuf1 wild-type growth is restored by administration, the growth deficiency of the cuf1 variant is due to impaired copper absorption.
본 발명에서, 성장이 결핍된 전사인자 변이체(24가지 변이체)를 2가지 그룹으로 분류하였다: (1) 온도-비의존성 성장 결핍 전사인자 변이체; (2) 온도-의존성 성장 결핍 전사인자 변이체. 첫 번째 전사인자 그룹(온도 비의존성)에는 BZP2, CUF1, HOB1, GAT5, FZC6 및 NRG1이 포함되어 있다(도 3). HXL1, CRX1, ATF1, ADA2, LIV4, ARO80, USV101, FZC31, MLN1, FZC30, FZC1, MIZ1, FZC46, APN2, GAT6, MBS2, SRE1, 및 ERT1을 포함하는 2차 그룹에 포함되는 전사인자의 결실은 고온(37℃~39℃)에서의 성장 결핍을 나타내었다(도 3). 이 중에서, UPR 신호전달경로 내 Ire1 키나아제의 하류에 있는 HXL1만이 숙주의 생리학적 온도에서 심각한 성장 결핍을 나타내었다. 반대로, MLN1 , MCM1 및 FZC46의 결실은 39℃에서 크립토코커스 네오포만스의 성장을 촉진시켰다. 종합하면, 이러한 결과는 다양한 전사인자(27가지 전사인자)들이 크립토코커스 네오포만스의 성장 및 열 내성에 대하여 양성 또는 음성적으로 조절한다는 것을 제시하였다.In the present invention, growth-deficient transcription factor variants (24 variants) were classified into two groups: (1) temperature-independent growth-deficient transcription factor variants; (2) Temperature-dependent growth-deficient transcription factor variants. The first group of transcription factors (temperature independent) included BZP2 , CUF1 , HOB1 , GAT5 , FZC6 and NRG1 (Fig. 3). The deletion of transcription factors included in the secondary group including HXL1 , CRX1 , ATF1 , ADA2 , LIV4 , ARO80 , USV101 , FZC31 , MLN1 , FZC30 , FZC1 , MIZ1 , FZC46 , APN2 , GAT6 , MBS2 , SRE1 , and ERT1 It showed a lack of growth at high temperature (37 ℃ ~ 39 ℃) (Figure 3). Among them, only HXL1 downstream of the Ire1 kinase in the UPR signaling pathway showed severe growth deficit at the physiological temperature of the host. In contrast, deletion of MLN1 , MCM1 and FZC46 promoted the growth of Cryptococcus neoformans at 39°C. Taken together, these results suggested that various transcription factors (27 transcription factors) positively or negatively regulate Cryptococcus neoformans growth and heat tolerance.
교배(mating)을Mating 조절하는 전사 인자 Regulating transcription factor
일반적인 환경에서, 크립토코커스 네오포만스는 주로 효모 형태로 존재하는데, 상반된 교배형 세포와 양성 분화를 진행하거나 같은 교배형 세포와 단성 분화를 진행하여 실 형태가 되고, 감염성 번식체인 담자 포자를 생산하게 된다. 이러한 발생학적 과정은 병원균의 유전적 다양성에 기여하게 된다(비특허문헌 17).In a general environment, Cryptococcus neoformans mainly exist in the form of yeast, and they undergo positive differentiation with opposite hybrid cells or unilateral differentiation with the same hybrid cells to form a thread, and produce basidiomycete spores, an infectious breeding cell. These embryological processes contribute to the genetic diversity of pathogens (Non-Patent Document 17).
교배 표현형을 분석하기 위해, 각각의 전사인자 변이체(혈청형 A MAT 균주)는 혈청형 A MATa 야생형 KN99a 균주(듀크대학교 Joeseph Heitman 연구실로부터 입수)와 공통배양함으로써 일방적인 교차교배를 구축하였다. 각각의 균주는 YPD 배지 내 30℃에서 16시간 동안 배양하였고, 같은 농도의 세포(107 세포/㎖)를 혼합하여 섞어서, V8 교배 배지(pH 5, 1L 기준: V8 juice 50 ㎖ (Campbell), KH2PO4 (Bioshop, PPM302) 0.5g, Agar (Bioshop, AGR001.500) 40g)에 스팟팅하였으며, 1 내지 2주간 상온에서 빛을 차단하여 배양하였다. 사상성 생육(filamentous growth)은 주마다 관찰하였고 스팟(SPOT) 통찰 디지털 카메라(Diagnostic Instrument Inc.)가 장착된 올림푸스 BX51 현미경을 사용하여 촬영하였다.To analyze the mating phenotype, each transcription factor variant (serum type A MAT strain) was co-cultured with serotype A MAT a wild type KN99a strain (obtained from Duke University's Joeseph Heitman laboratory) to construct unilateral crosscrossing. Each strain was cultured for 16 hours at 30° C. in YPD medium, and cells of the same concentration (10 7 cells/ml) were mixed and mixed, and V8 mating medium (
페로몬 유전자의 발현을 관찰하기 위한 세포-융합 시험 및 노던 블롯 분석을 실시하였다. 세포-융합 시험에서, NatR마커를 포함하는 MAT 전사인자 변이체 또는 대조군 균주(YSB119, 듀크대학교 Joeseph Heitman 연구실로부터 입수) 및 네오마이신-내성(Neor)마커를 포함하는 MAT a 대조군 균주(YSB121, 듀크대학교 Joeseph Heitman 연구실로부터 입수)는 각각 액체 YPD 배지 내 30℃에서 16시간 동안 배양하였고, 세포의 농도는 증류수를 사용하여 107 세포/㎖로 맞추었다. 각각의 MAT 균주 및 MATa 균주는 동일한 부피로 혼합하여 V8 배지에 스팟팅하였고 상온에서 24시간 동안 빛을 차단하여 배양하였다. 이후에, 세포를 긁어 모아서 증류수 1㎖에 재부유시켰고, 노르세트로신(100 ㎍/㎖) 및 G418(50 ㎍/㎖, Geneticin, Life technologies)을 모두 포함하는 YPD 배지 플레이트 상에 펼쳐놓았다. 이후에, 상기 플레이트는 30℃에서 추가 배양하였고 각각의 플레이트 상의 콜로니 수를 측정하였다. 페로몬 유전자 발현 모니터링에서, MAT 균주 및 KN99a 균주는 동일한 세포 농도(108 세포/㎖)로 혼합하여 V8 배지에 펼쳐놓고 상온에서 18시간 내지 24시간 동안 빛을 차단하여 배양하였다. 이후에 세포는 V8 배지로부터 긁어내어 4℃에서 펠릿 형태로 제조하였고, 액체질소에서 동결시켰으며 하룻밤 동안 동결 건조시켰다. 전체 RNA는 선행 문헌에 기재된 프로토콜에 따라 트라이졸(Trizol agent)을 사용하여 분리하였다. RNA를 전기영동한 후 멤브레인으로 이동시켰고, 멤브레인은 프라이머 B1894(5'- TTTTACGCTTTTTGCAGATTCCGCCAAA -3'), B195(5'- GACCACTGTTTCTTTCGTTCT -3') 및 JEC21 게놈 DNA(JEC21 균주에서 게놈 DNA를 추출)를 사용하여 증폭시켜 제조된 교배 페로몬 유전자(MF1)-특이적 탐침과의 혼성화를 실시하였다.Cell-fusion tests and Northern blot analysis were performed to observe the expression of the pheromone gene. In the cell-fusion test, the MAT transcription factor variant or control strain containing the Nat R marker (YSB119, obtained from the Joeseph Heitman laboratory at Duke University) and the MAT a control strain containing the neomycin-resistant (Neo r ) marker (YSB121, Duke University (obtained from Joeseph Heitman's laboratory) was cultured in liquid YPD medium at 30° C. for 16 hours, and the concentration of cells was adjusted to 10 7 cells/ml using distilled water. Each MAT strain and MAT a strain were mixed in the same volume, spotted on V8 medium, and cultured by blocking light for 24 hours at room temperature. Thereafter, the cells were scraped and resuspended in 1 ml of distilled water, and spread on a YPD medium plate containing both norsetrosine (100 μg/ml) and G418 (50 μg/ml, Geneticin, Life technologies). Thereafter, the plates were further incubated at 30° C. and the number of colonies on each plate was measured. In monitoring pheromone gene expression, the MAT strain and the KN99 a strain were mixed at the same cell concentration (10 8 cells/ml), spread on a V8 medium, and cultured at room temperature by blocking light for 18 to 24 hours. Thereafter, the cells were scraped from the V8 medium and prepared in the form of pellets at 4° C., frozen in liquid nitrogen, and freeze-dried overnight. Total RNA was isolated using a Trizol agent according to the protocol described in the prior literature. RNA was transferred to the membrane after electrophoresis, and the membrane used primers B1894 (5'- TTTTACGCTTTTTGCAGATTCCGCCAAA -3'), B195 (5'- GACCACTGTTTCTTTCGTTCT -3') and JEC21 genomic DNA (genomic DNA extracted from the JEC21 strain). Hybridization with a cross pheromone gene (MF1)-specific probe prepared by amplification was performed.
교배 표현형을 분석하기 위해서, 혈청형 MAT a KN99a 균주와 각각의 전사인자 변이체를 공통-배양함으로써 일방적인 교차 교배를 구축하였다. 본 발명에서의 신규-교배 조절 전사인자 중에서, BZP2, USV101, FZC1 및 ZAP104의 결실은 교배(특히, 단성 교배)를 크게 감소시키지만, HLH1, HAP2 및 GAT1의 결실은 교배 효율을 크게 향상시켰다(도 6a). 섬사(filamentation) 과정에 선행하는 세포 융합 및 페로몬 생산의 효율을 측정하여 상기 전사인자가 관련되어 있는 교배 단계를 검증하였다. bzp2, usv101, fzc1 및 zap104 변이체는 MAT a 대조군 균주와의 세포 융합이 나타나지 않았고(도 6b), 교배 시 페로몬 유전자(MF1)의 발현이 나타나지 않았다(도 4c). 이러한 결과는 Bzp2, Usv101, Fzc1 및 Zap104 전사인자가 페로몬 유전자 발현을 촉진하여 세포 융합을 증가시킨다는 것을 제시하였다. 반대로, hlh1, hap2, 및 gat1 변이체 내에서 세포-융합 효율은 2-3 배 증가하였고(도 6b), 페로몬 유전자 발현은 크게 증가하였다(도 6c). 결실되면 교배를 촉진시키는 SKN7은 페로몬 유전자 발현 및 세포 융합에 필수적인데(도 6b 및 도 6c), 이는 SKN7이 교배의 후반부 단계에 관련되어 있음을 제시하였다(도 6d). 본 발명에 따른 분석에서 34가지 전사인자가 교배에 관련되어 있음을 제시하였다(도 7a- 양성조절인자, 도 7b-음성조절인자).In order to analyze the mating phenotype, unilateral cross mating was constructed by co-culturing the serotype MAT a KN99 a strain and each of the transcription factor variants. Among the novel-cross-regulating transcription factors in the present invention, deletion of BZP2 , USV101 , FZC1 and ZAP104 greatly reduced mating (particularly, single mating), but deletion of HLH1 , HAP2 and GAT1 greatly improved mating efficiency (Fig. 6a). By measuring the efficiency of cell fusion and pheromone production prior to the filamentation process, the mating step in which the transcription factor is related was verified. The bzp2 , usv101 , fzc1 and zap104 mutants did not show cell fusion with the MAT a control strain (Fig. 6b), and did not show the expression of the pheromone gene (MF1) during mating (Fig. 4c). These results suggested that Bzp2, Usv101, Fzc1, and Zap104 transcription factors promote pheromone gene expression to increase cell fusion. Conversely, in the hlh1 , hap2 , and gat1 mutants, the cell-fusion efficiency increased 2-3 times (FIG. 6B ), and the pheromone gene expression was significantly increased (FIG. 6C ). When deleted,
독성 인자 생산을 조절하는 전사 인자Transcription factors that regulate the production of toxic factors
숙주 내 생존 및 증식에 있어서, 크립토코커스 네오포만스는 캡슐 및 멜라닌을 포함하는 다양한 독성인자를 갖추고 있다. 캡슐은 숙주 식균세포에 의한 식균작용으로부터 세포를 보호하는 클루코로노크실로만난(glucuronoxylomannan) 또는 갈락토실로만난(galactoxylomannan)에 기반한 다당류이다(비특허문헌 18). 멜라닌은 폴리페놀 복합체로 만들어진 검-갈색 색소로서, 세포에 항식균능 및 항산화능을 제공한다(비특허문헌 19).For survival and propagation in the host, Cryptococcus neoformans possess a variety of virulence factors including capsules and melanin. The capsule is a polysaccharide based on glucuronoxylomannan or galactoxylomannan that protects cells from phagocytosis by host phagocytic cells (Non-Patent Document 18). Melanin is a black-brown pigment made of a polyphenol complex, and provides anti-phagocytosis and antioxidant activity to cells (Non-Patent Document 19).
5.1 캡슐 생산5.1 Capsule production
캡슐 생산은 선행 문헌에 기재된 바와 같이 정성 및 정량적 방법에 의해 측정하였다(비특허문헌 20 및 비특허문헌 21). 세포는 액체 YPD 배지 내 30℃에서 16시간 동안 성장시켰고, DME(Dulbeccos modified Eagle) 고체 배지(Gibco, #12100-046)로 스팟팅하였으며, 이후에 2일간 37℃에서 배양하였다. 배양 후에, 세포는 DME 고체 배지로부터 긁어내어 PBS(Phosphate-Buffed Saline)으로 세척하였다. 캡슐은 인디아 잉크(Bactidrop; Remel)로 염색하고 스팟 관찰 디지털 카메라(Diagnostic Instrument Inc.)를 갖춘 올림푸스 BX51 현미경을 사용하여 시각화함으로써 캡슐 생산을 정성 측정하였다. 정성 분석을 위해, DME 고체 배지로부터 수집된 세포는 10% 포르말린으로 고정시켰고, 같은 수의 세포(2.5×107 세포/㎖)를 헤마토크릿 모세관에 로딩하였다. 상기 헤마토크릿 모세관은 수직으로 배치하여 세포가 10일 동안 중력에 의해 압축되도록 하였다. 압축된 세포 부피 비율은 전체 부피 상(세포+배지)의 길이에 대한 압축된 세포 부피 상의 길이 비율을 계산하여 측정하였다. 각각 변이체의 상대적인 압축 세포 부피는 야생형 압축 세포 부피 비율에 대한 변이체 압축 세포 부피 비의 비율을 계산하여 측정하였다. Capsule production was measured by qualitative and quantitative methods as described in prior literature (
2 가지 또는 2 가지 이상의 독립적인 균주를 이용한 3회 반복 실험을 실시하였다. 상대적으로 압축된 세포 부피에서의 통계적 차이는 Prism 6(Graph pad software)를 이용한 본페로니(Bonferroni) 다중 비교 테스트를 포함하는 일월변량분석(one way ANOVA)에 의해 측정하였다.The experiment was repeated three times using two or two or more independent strains. Statistical differences in the relatively compressed cell volume were measured by one way ANOVA including a Bonferroni multiple comparison test using Prism 6 (Graph pad software).
본 발명에서 캡슐 생산에 49가지 전사인자가 관련되어 있음을 확인하였다(도 8a, 8b-음성조절인자 29가지, 8c-양성조절인자 20가지). 기존에 알려진 캡슐-조절 전사인자인 Atf1(비특허문헌 10), Mbs1(비특허문헌 11), Gat201(비특허문헌 12) 및 Ada2(비특허문헌 22)를 포함하고 있다. 이러한 캡슐 조절 전사인자에 부가하여, 본 발명자는 여러 가지 신규 캡슐-조절 전사인자를 확인하였다. 기존에 알려진 gat201Δ 및 ada2Δ 변이체에서 관찰된 결과에 비해 zap104Δ, bap1Δ 및 rds2Δ 변이체는 캡슐 생산이 크게 감소하였다(도 8a). 따라서, Ada2, Gat201, Zap104, Bap1 및 Rds2는 다른 15가지 양성 조절인자와 함께 캡슐 생산에서의 주요한 양성 조절인자로 예측되었다. 반대로, HOB7, CLR3 및 FZC51의 결실은 캡슐 생산을 크게 향상시켰는데(도 8a), 이는 다른 26가지 음성 조절인자와 함께 캡슐 생산의 주요한 음성 조절인자로 예측되었다.In the present invention, it was confirmed that 49 transcription factors were involved in capsule production (FIG. 8a, 8b-
5.2 멜라닌 생산 측정 및 5.2 Measurement of melanin production and LAC1LAC1 발현 분석 Expression analysis
각각의 전사인자 변이체 균주 및 야생형 균주는 액체 YPD 배지 내 30℃에서 하룻밤 동안 배양하여, 0.1% 또는 0.3% 글루코스를 포함하는 니거 종자(Niger seed) 아가 배지(Kaytee)에 스팟팅하였고 이후에 37℃에서 배양하였는데 멜라닌 생산을 관찰하여 매일 촬영하여 멜라닌 생산을 측정하였다. 본 발명에서는 멜라닌 생산에 관여하는 27가지 전사인자를 확인하였다(도 8d, 9a- 양성조절인자 11가지, 9b- 음성조절인자 16가지). Cuf1, Ste12, Mbs1, Skn7, 및 Atf1을 포함하는 소수의 전사인자는 기존에 공개된 전사인자이다(비특허문헌 10, 비특허문헌 11, 비특허문헌 23, 비특허문헌 24 및 비특허문헌 25). 이러한 결과에 부가하여, fzc8Δ, hob1Δ 및 bzp4Δ 변이체가 멜라닌 생산이 크게 감소하였음을 확인하였는데, cuf1Δ 변이체의 결과와 유사하였다(도 9a).Each of the transcription factor variant strains and wild-type strains were cultured overnight at 30° C. in liquid YPD medium and spotted on Niger seed agar medium (Kaytee) containing 0.1% or 0.3% glucose, and then 37° C. It was cultured in, and the melanin production was observed and photographed every day to measure melanin production. In the present invention, 27 transcription factors involved in melanin production were identified (Fig. 8d, 9a-
또한, 본 발명에서 멜라닌 합성에 관련된 주요 락카제(laccase)인 LAC1 발현과 전사인자 사이의 관련성을 검증하였다. 우선, 노던 블롯 분석을 실시하여 LAC1 유도를 측정하였다. 야생형 및 각각의 전사인자 변이체는 YPD 배지 내 30℃에서 16시간 동안 배양하였고, 배양된 세포는 600 nm에서의 광학 밀도(OD600)를 0.15로 맞추어서 신선한 YPD 액체 배지 150ml로 접종하였다. 세포 배양액은 OD600이 대략 0.6이 될 때까지 30℃에서 추가로 배양하였다. 제로 타임 시료에 대하여, 150 ㎖ 중 50 ㎖을 표본 채취하였고, 남은 100 ㎖ 배양액은 원심분리에 의해 펠릿을 제조하였다. 상층액을 제거한 후, 세포는 글루코스 없는 YNB 액체 배지(Becton, Dickison and company, #291940)를 사용하여 재부유하였다. 배양 중에, 50 ㎖ 배양액은 1시간 및 2시간에 표본 채취하였다. 전체 RNA는 각각의 표본으로부터 추출하였고, H99 게놈 DNA, 프라이머 B3662(5'-CTTTCAATCGTCCAAGCG-3') 및 프라이머 B3663(5'-CCCCAGTTATCCAAAAAGTC-3')을 이용한 이용한 PCR에 의해 증폭시킨 LAC1 특이적 탐침을 사용하여 노던 블롯을 실시하였다. 이에 의해, 본 발명에서는 Hob1 및 Fzc8이 글루코스-결핍 조건에서 멜라닌 합성에 관련된 주요 락카제(laccase)인 LAC1의 발현을 촉진하는 것을 확인하였다(비특허문헌 26). Bzp4 및 Cuf1은 LAC1 발현에 직접적으로 관련되어 있지 않았다(도 8e). 반대로, Hlh1은 LAC1 발현을 음성적으로 조절하였지만 HLH1, HLH2, BAP1 및 FZC1의 결실은 멜라닌 생산을 크게 증가시켰다(도 8e). 따라서, 본 발명에서 크립토코커스 네오포만스 내 LAC1의 신규 양성 조절인자(Hob1 및 Fzc45) 및 음성 조절인자(Hlh1)를 확인하였다. In addition, in the present invention, the relationship between the expression of LAC1 , a major laccase related to melanin synthesis, and a transcription factor was verified. First, Northern blot analysis was performed to measure LAC1 induction. Wild-type and each of the transcription factor variants were incubated for 16 hours at 30° C. in YPD medium, and the cultured cells were inoculated with 150 ml of fresh YPD liquid medium by adjusting the optical density (OD 600) at 600 nm to 0.15. The cell culture solution was further incubated at 30° C. until an OD 600 reached approximately 0.6. For the zero time sample, 50 ml of 150 ml was sampled, and the remaining 100 ml culture solution was centrifuged to prepare a pellet. After removing the supernatant, the cells were resuspended using YNB liquid medium without glucose (Becton, Dickison and company, #291940). During cultivation, 50 ml culture was sampled at 1 and 2 hours. Total RNA was extracted from each sample, and a LAC1-specific probe amplified by PCR using H99 genomic DNA, primer B3662 (5'-CTTTCAATCGTCCAAGCG-3') and primer B3663 (5'-CCCCAGTTATCCAAAAAGTC-3') was used. Northern blot was carried out. Accordingly, in the present invention, it was confirmed that Hob1 and Fzc8 promote the expression of LAC1 , which is a major laccase related to melanin synthesis under glucose-deficient conditions (Non-Patent Document 26). Bzp4 and Cuf1 were not directly related to LAC1 expression (FIG. 8E ). Conversely, Hlh1 negatively regulated LAC1 expression, but deletion of HLH1 , HLH2 , BAP1 and FZC1 significantly increased melanin production (FIG. 8E). Therefore, in the present invention, novel positive regulators (Hob1 and Fzc45) and negative regulators (Hlh1) of LAC1 in Cryptococcus neoformans were identified.
5.3 우레아제(5.3 Urease ( UreaseUrease ) 생산) production
크립토코커스 네오포만스의 독성에 관련된 주요 인자는 캡슐, 멜라닌 외에 유레아제가 있는데, 이는 요소(Urea)를 암모니아로 전환시켜 질소원을 제공하는 니켈-의존성 단백질 복합체이다(Ure1, Ure4, Ure6, 및 Ure7). The main factors involved in the toxicity of Cryptococcus neoformans are capsules and melanin, as well as urease, which is a nickel-dependent protein complex that provides a nitrogen source by converting urea to ammonia (Ure1, Ure4, Ure6, and Ure7). .
야생형 균주 및 전사인자 변이체 균주를 액체 YPD 배지 내 30℃에서 16시간(하룻밤) 동안 배양하였고, 증류수로 세척한 후 세포 밀도를 1×107 세포/㎖로 맞추었다. 이후에 5 ㎕ 세포(5×104 세포)는 크리스텐젠 아가 배지에 스팟팅하였고, 30℃에서 7일 내지 10일간 배양하였으며, 배양 중에 세포를 촬영하였다. The wild-type strain and the transcription factor variant strain were cultured in liquid YPD medium at 30° C. for 16 hours (overnight), washed with distilled water, and then the cell density was adjusted to 1×10 7 cells/ml. Thereafter, 5 μl cells (5×10 4 cells) were spotted on Christengen agar medium, cultured at 30° C. for 7 to 10 days, and cells were photographed during the culture.
본 발명에서 우레아제 생산을 음성 조절(4가지, 도 10a) 또는 양성 조절(15가지, 도 10b)하는 19가지 전사인자를 확인하였다. In the present invention, 19 transcription factors that negatively regulate urease production (4 types, Fig. 10a) or positive control (15 types, Fig. 10b) were identified.
크립토코커스Cryptococcus 네오포만스Neoformance 독성에 영향을 주는 전사인자 Transcription factors affecting toxicity
크립토코커스 네오포만스의 병원성에 필요한 전사인자의 검증은 항진균제 및 치료 방법의 개발에 매우 중요한 과정이다. 본 발명에서 2가지 대량 독성 실험을 실시하였다: (1) 곤충 애벌레 모델 시스템 벌집나방(Galleria mellonella) 내에서 수행한 시험; 및 (2) 쥐과 동물 호흡 모델에서의 서명-표지 변이(STM)-기반의 진균 실험. 곤충 숙주 및 포유류 숙주 모델을 사용한 실험은 크립토코커스 네오포만스 뿐만 아니라 다른 진균류에 대한 대량-독성/감염 시험에서 널리 적용되고 있다(비특허문헌 12).Verification of transcription factors necessary for the pathogenicity of Cryptococcus neoformans is a very important process in the development of antifungal agents and treatment methods. Two mass toxicity experiments were conducted in the present invention: (1) a test performed in the insect larvae model system honeycomb moth (Galleria mellonella); And (2) signature-label mutation (STM)-based fungal experiments in a murine respiratory model. Experiments using insect host and mammalian host models have been widely applied in mass-toxicity/infection tests against Cryptococcus neoformans as well as other fungi (Non-Patent Document 12).
6.1 곤충 시험6.1 Insect test
곤충 기반 독성 시험은 기존 방법(비특허문헌 21)을 일부 변형하여 실시하였다. 선적일(vanderhorst Inc., St Marys, OH, USA)로부터 7일 이내에 도착한 최종 영 유충기 단계의 15수의 벌집나방(Galleria mellonella) 애벌레(체중:250±50 ㎎)를 각각의 그룹에 무작위로 분류한 후 벌집나방 살상 시험을 실시하였다. 각각의 크립토코커스 네오포만스 균주는 YPD 배지 내 30℃에서 하룻밤 동안 배양한 후, PBS로 3회 세척하고 나서 PBS에 재부유시켜서 혈구계산기를 이용한 세포 수 계산에 사용하였다. 본 발명자들은 10 ㎕ 크기의 바늘 및 반복적인 분배기(PB600-1, Hamilton)을 갖춘 100 ㎕ 해밀튼 주사기를 사용하여 애벌레의 최종 전각에 4,000개의 크립토코커스 네오포만스 세포를 2차로 접종하였다. 비-감염 대조군에는 PBS를 주입하였다. 주입 후에, 애벌레는 가습된 플라스틱 컨테이너 내 페트리 디쉬에서 배양하였고 매일 관찰하였다. 감염된 애벌레는 37℃에서 배양하였고, 매일 관찰하였다. 애벌레는 만졌을 때 움직임이 없으면 죽은 것으로 간주하였다. 실험 중에 번데기로 변이된 애벌레는 통계 분석 대상에서 제외시켰다. 생존 곡선은 Prism 6(GraphPad)를 사용하여 작성하였고, 로그-순위(Mantel-Cox) 테스트를 사용하여 통계적으로 분석하였다. 우선, 각각의 전사인자 유전자(전체 155가지)에 대한 단일 변이체 균주의 생존 곡선을 관찰하였고, 이를 야생형 균주의 생존 곡선과 통계적으로 비교하였다. 유전자 결실에 의해 통계적으로 유의한 수준(P<0.05; 로그-순위 테스트)의 독성 감소 또는 향상을 나타내는 전사인자 변이체에 대하여 2차의 독립된 균주를 검사하였다.Insect-based toxicity test was conducted with some modifications of the existing method (Non-Patent Document 21). 15 number of beehive moths (Galleria ) of the last juvenile stage, arriving within 7 days of the date of shipment (vanderhorst Inc., St Marys, OH, USA) mellonella ) larvae (weight: 250±50 mg) were randomly classified into each group, and a honeycomb moth killing test was conducted. Each Cryptococcus neoformans strain was cultured overnight at 30° C. in YPD medium, washed three times with PBS, and then resuspended in PBS, and used for counting cells using a hemocytometer. We used a 100 μl Hamilton syringe equipped with a 10 μl size needle and a repeat dispenser (PB600-1, Hamilton) to inoculate 4,000 Cryptococcus neoformans cells secondarily into the final anterior leg of the larvae. PBS was injected into the non-infected control group. After injection, larvae were cultured in Petri dishes in a humidified plastic container and observed daily. Infected larvae were cultured at 37°C and observed daily. Caterpillars were considered dead if they were not moving when touched. The larvae transformed into pupa during the experiment were excluded from the statistical analysis. Survival curves were created using Prism 6 (GraphPad) and statistically analyzed using log-rank (Mantel-Cox) test. First, the survival curve of the single variant strain for each transcription factor gene (total 155 types) was observed, and this was statistically compared with the survival curve of the wild-type strain. Secondary independent strains were examined for transcription factor variants showing statistically significant levels (P<0.05; log-rank test) reduction or enhancement of toxicity by gene deletion.
각각의 패널은 각각의 전사인자에 대하여 2가지 독립적인 변이체에 대한 생존 결과를 포함하고 있다(도 11). 검증된 변이체는 전신 크립토코카스증의 쥐과 모델 내 독성 감소를 나타내는 것으로 기존에 알려진 9가지 전사인자 변이체(hxl1, ada2, sre1, nrg1, bwc2, crz1, pdr802, gat201 및 gat204)를 포함하였다(비특허문헌 9, 비특허문헌 12, 비특허문헌 22, 비특허문헌 27, 비특허문헌 28, 비특허문헌 29, 비특허문헌 30, 비특허문헌 31 및 비특허문헌 32). 이러한 결과는 곤충 모델 및 쥐과 동물 모델이 크립토코커스 네오포만스 병원성에서 깊은 관련성이 있음을 제시하였다. 알려진 전사인자의 변이 외에도, HOB1, BZP2, USV101, BAP1, ZFC2, FZC1, FZC50, 및 FZC31의 변이는 크립토코커스 네오포만스의 독성을 크게 감소시켰다(도 11). 본 발명에서는 곤충 숙주 모델을 사용하여 크립토코커스 네오포만스의 독성에 관련된 17 가지 전사인자 유전자를 검증하였다(도 11).Each panel contains survival results for two independent variants for each transcription factor (Fig. 11). Validated variants included nine transcription factor variants (hxl1 , ada2 , sre1 , nrg1 , bwc2 , crz1 , pdr802 , gat201 and gat204 ) previously known to exhibit reduced toxicity in a murine model of systemic cryptococcosis (non-patented
6.2 동물 시험6.2 Animal testing
동물 관리 및 모든 실험은 연세대학교 실험동물 위원회(IACUC, Institutional Animal Care and Use Committe)의 지침에 따라 실시하였다. 연세대학교 IACUC는 척추동물 연구에 관한 모든 사항을 승인하였다.Animal care and all experiments were conducted according to the guidelines of Yonsei University's Institutional Animal Care and Use Committe (IACUC). Yonsei University IACUC approved all matters related to vertebrate animal research.
서명-표지 변이(STM)에 기반하는 마우스 감염 연구에서, 44가지 다른 서명 표지(표 1)로 표지된 전사인자 균주들은 YPD 배지 내 30℃에서 배양하였고, PBS로 3회 세척한 후 풀을 형성하였다; 혈구 계산기를 사용하여 세포를 계산한 후 각각의 균주에 대하여 같은 수의 세포를 사용하였다. STM#282 및 STM#169 서열로 표지된 ste50 및 ire1 변이체는 각각 독성 대조군 및 비독성 대조군 균주로 사용하였다(비특허문헌 9 및 비특허문헌 33). 따라서, 동일한 서명 표지를 나타내는 전사인자(TF)의 수는 4 미만이다. 본 발명에서는 STM에 기반한 4가지의 독성 시험을 실시하였다. 인풋 전사인자 게놈 DNA 라이브러리를 획득하기 위해, 풀에 등록된 전사인자 변이체는 10배로 연속적으로 희석하여 YPD 배지에 플레이팅하고, 3일간 30℃에서 배양시켰고, 긁어내어 모은 후에 게놈 DNA 추출에 사용하였다. 아웃풋 전사인자 게놈 DNA 라이브러리는 하기와 같이 실시하였다. 아베르틴(avertin, 2,2,2-트리브로모에탄올)의 복강내 주사에 의해 마취시킨 7주령 암컷 A/Jcr 마우스(Jacson Laboratory)는 등록된 전사인자 변이체 5×105세포(50 ㎕ 부피)를 비강내 흡입에 의해 감염시켰고 감염 후 15일에 아베르틴을 초과투여하여 희생시켰다. 각각의 검사에 대하여, 본 발명에서는 마우스 5두를 사용하였다. 각각의 전사인자에 대한 2가지 독립된 변이체는 구분된 STM 세트 시험에서 검사하였다. 마우스 폐는 절단한 후 PBS를 사용하여 균질화하였다. 각각의 폐-조직 용해물은 100 ㎍/㎖ 클로로암페니콜을 포함하는 YPD 배지에 펼친 후 3일 동안 30℃에서 배양하였다. 배양 후 긁어내어 모아서 아웃풋 게놈 DNA를 추출하였다. 인풋 및 아웃풋 게놈 DNA는 CTAB 방법에 따라 추출하였다(비특허문헌 21). 표 S1에 기재된 다양한 표지-특이적 프라이머 및 MyiQ2 리얼-타임 PCR 탐지 시스템(Bio-Rad)을 사용하여 정량적 PCR 분석을 실시하였다. STM 수치(Log2[아웃풋/인풋])는 선행문헌에 기재된 방식에 따라 계산하였다(비특허문헌 12, 비특허문헌 34).In a mouse infection study based on signature-label mutation (STM), transcription factor strains labeled with 44 different signature markers (Table 1) were cultured at 30° C. in YPD medium, washed 3 times with PBS, and then formed a pool. Did; After counting the cells using a hemocytometer, the same number of cells was used for each strain. The ste50 and ire1 variants labeled with the
6.3 동물 시험 결과6.3 Animal test results
STM-기반의 쥐과동물-숙주 모델을 사용하여, 40가지 독성 유전자를 검증하였다(도 11d). 각각의 변이체에 대한 STM 수치는 희생시킨 마우스 폐 내에서 정량 PCR 수치(마우스 3두에 대한 평균 수치)=Log2(아웃풋/인풋)에 기반하여 계산하였다. 모든 실험 세트 중에서, 비독성 대조군 균주인 ire1Δ 변이체는 매우 낮은 STM 수치(-7.03±1.99)를 나타내었는데, 반면 독성 대조군 균주인 ste50Δ 변이체는 0.11±1.13의 STM 수치를 나타내었다. STM 수치 결과를 뒷받침하기 위해, 본 발명에서 확인된 40가지 유전자 중 11가지 유전자는 독성에 관련된 것으로 이미 알려져 있다(비특허문헌 9, 비특허문헌 12, 비특허문헌 29 및 비특허문헌 30). gat201Δ 변이체 및 pdr802Δ 변이체는 기존 문헌(비특허문헌 12)에 비해 크게 감소된 STM 수치(각각 -11.125 및 -7.212)를 나타내었다. 유사하게, zap104Δ 변이체 및 liv1Δ 변이체의 STM 수치 또한 감소하였다(각각 -5.528 및 -3.875). 반면, zap103Δ 변이체는 기존 문헌(비특허문헌 12)에 기재된 바와 같이 독성이 매우 높았다(STM 수치=2.51). 부가적으로, hxl1Δ 변이체, nrg1Δ 변이체, bwc2Δ 변이체 역시 크게 감소된 STM 수치를 나타내었다. STM 분석에 의해 확인된 40가지 전사인자 중 11가지 전사인자(GAT201, PDR802, HXL1, BWC2, NRG1, FZC1, HOB1, USV101, ZFC2, SRE1 및 FZC31)는 곤충 모델을 사용하여 재확인되었다. 곤충 모델을 이용한 독성 시험 자료는 피어슨 상관계수(PCC, Pearson Correlation Coefficient) 분석에 기반한 쥐과 동물 모델로부터의 STM-기반 감염성 자료와 통계적으로 연관성이 매우 높았다(도 12). STM-기반의 쥐과 동물 모델만을 사용하여 스크리닝한 26가지 신규 독성 관련 전사인자들의 변이체 중에서, fzc31Δ 변이체 및 ddt1Δ 변이체는 크게 감소된 STM 수치를 나타내었다(각각 -4.328 및 -4.832). 본 발명에 따른 발형체(phenome) 데이터베이스는 fzc31Δ 변이체가 삼투압, 산화 및 세포 막 스트레스에 대하여 증가된 감수성을 나타내었는데, 이러한 감수성이 모여서 독성에 영향을 주는 것으로 예측되었다. 반대로, ddt1Δ 변이체에서 관찰된 하나의 주목할 만한 표현형은 약한 DTT(dithiothreitol) 감수성인데, 다른 DTT-반응성 전사인자 변이체들이 야생형 균주만큼 독성이 있으므로, ddt1Δ 변이는 폐에서의 변이체 생존 감소에 연관되지 않은 것으로 예측되었다.Using the STM-based murine-host model, 40 virulence genes were verified (FIG. 11D). STM values for each variant were calculated based on quantitative PCR values (average values for 3 mice) = Log 2 (output/input) in the sacrificed mouse lung. Among all experimental sets, the non-toxic control strain, the ire1Δ variant, exhibited very low STM values (-7.03±1.99), while the toxic control strain, the ste50Δ variant, exhibited an STM value of 0.11±1.13. In order to support the STM numerical results, 11 of the 40 genes identified in the present invention are already known to be related to toxicity (Non-Patent
항균제 내성 및 감수성 검사Antimicrobial resistance and susceptibility test
*크립토코커스증의 치료에 암포테리신 B(AmpB), 플루사이토신(5-FC) 또는 이의 조합, 및 플루코나졸(FCZ)이 널리 사용되고 있다(비특허문헌 2). 하지만, 상기 약제의 고유한 독성 부작용 외에도, 항진균제-내성 크립토코커스 균주의 연속적인 발생은 심각한 임상 문제를 유발하였다(비특허문헌 35). 본 발명에서는 크립토코커스 네오포만스 TFKO 군집 내 균주들 간의 항진균제 감수성 변화를 관찰하여 항진균제 내성에 관련된 전사인자를 검증하였다.* Amphotericin B (AmpB), flucytosine (5-FC), or a combination thereof, and fluconazole (FCZ) are widely used in the treatment of cryptococcus (Non-Patent Document 2). However, in addition to the inherent toxic side effects of the drug, the continuous occurrence of antifungal-resistant Cryptococcus strains caused serious clinical problems (Non-Patent Document 35). In the present invention, the transcription factor related to antifungal resistance was verified by observing the change in antifungal agent sensitivity between strains in the Cryptococcus neoformans TFKO community.
세포는 액체 YPD 배지 내 30℃에서 16시간 동안 배양하였고, 10배 단위로 순차적 희석을 실시하였으며(1 내지 104배 희석), 항균제(fludiooxonil, fluconazole, amphotericin B, flucytosine)의 지정된 농도를 포함하는 YPD 배지 상에 세포를 스팟팅하였다(spotted). 세포는 30℃에서 배양하였고 2일 내지 5일간 촬영하였다.Cells were cultured for 16 hours at 30° C. in liquid YPD medium, and sequentially diluted in 10-fold increments (1 to 10 4- fold dilution), containing a specified concentration of antimicrobial agents (fludiooxonil, fluconazole, amphotericin B, flucytosine). Cells were spotted on YPD medium. Cells were cultured at 30° C. and photographed for 2 to 5 days.
다수의 전사인자가 항진균 내성에 관련되어 있음을 확인하였는데(도 13), 이는 크립토코커스 균주가 다양한 방법으로 항진균제에 적응할 수 있다는 것을 암시하였다. 플루코나졸(FCZ)에 대한 반응에서, 35.5%(55/155)의 전사인자가 향상된 감수성(35가지 전사인자) 또는 내성(20가지 전사인자)을 나타내었는데, 이는 아졸계 항진균제에 대한 내성이 다양한 전사인자의 조절에 의해 즉각적으로 발생함을 암시하는 결과이다. 하지만, 암포테리신(AmpB)에 대한 반응에서 56가지 유전자의 변이체는 다양한 약제 감수성을 나타내었는데 47가지 전사인자 변이체는 감수성이 증가하였고 8가지 전사인자 변이체는 약제에 대한 내성 증가를 나타내었다(도 13). 이러한 자료는 아졸 내성에 비해 폴리엔 약제에 대한 내성이 거의 관찰되지 않는 임상 결과를 뒷받침하였다. 추가적으로, 5-FC는 균주 내 약제 내성을 빠르게 유발한다고 알려져 있다(비특허문헌 36). 이러한 결과를 뒷받침하는 본 발명에서는 27가지 전사인자가 플루사이토신 내성을 차별적으로 조절하는 것을 확인하였다(도 13). It was confirmed that a number of transcription factors were involved in antifungal resistance (FIG. 13 ), which suggested that the Cryptococcus strain could adapt to antifungal agents in various ways. In response to fluconazole (FCZ), 35.5% (55/155) of transcription factors showed improved susceptibility (35 transcription factors) or resistance (20 transcription factors). It is a result suggesting that it occurs immediately by the regulation of factors. However, in response to amphotericin (AmpB), 56 gene variants exhibited various drug sensitivity, 47 transcription factor variants increased sensitivity, and 8 transcription factor variants increased drug resistance (Fig. 13). These data supported the clinical results in which little resistance to polyene drugs was observed compared to azole resistance. Additionally, 5-FC is known to rapidly induce drug resistance within the strain (Non-Patent Document 36). In the present invention supporting these results, it was confirmed that 27 kinds of transcription factors differentially regulate flucytosine resistance (FIG. 13).
본 발명에서는 일부 전사인자의 결실이 아졸 및 폴리엔 약제에 대한 감수성을 음성 조절한다는 것을 확인하였는데(도 13 및 도 14a), 이는 상기 전사인자들이 ERG11 발현 및 스테롤 생합성을 직접 조절하여 폴리엔 결합 능력에 영향을 주기 때문이다. 상기 전사인자 중 2가지는 Erg11 조절인자로 이미 알려져 있다. Sre1은 스테롤의 주요 조절인자로서, 크립토코커스 네오포만스 내 스테롤 조절 인자-결합 단백질(SREBP)의 일부분인 Scp1과 복합체를 형성하였다(비특허문헌 27 및 비특허문헌 28). Mbs1은 기본적인 ERG11 발현을 음성 조절하므로 크립토코커스 네오포만스 내 Mbs1 결실은 아졸 내성을 증가시키지만 폴리엔 내성은 감소시킨다(비특허문헌 11). 스테롤-충분 조건 및 스테롤-결핍 조건에서 전사인자 변이체 내 ERG11 발현을 측정하여 ERG11 발현에 관련된 다른 전사인자를 검사하였다(도 14b). ERG2, ERG3, ERG5, ERG11, 및 ERG25의 발현을 분석하기 위해, 야생형, sre1, 및 hob1을 액체 YPD 배지에서 하룻밤 동안 배양하였다. 배양액은 다시 신선한 30℃ YPD 배지 100 ㎖에 접종하였고, 배양물의 OD600수치가 대략 1.0이 될 때까지 배양하였다. 제로-타임 시료를 준비하기 위해, 세포 배양액 50 ㎖을 표본 채취하였고, 남은 배양물은 플루코나졸(최종 농도: 10 ㎍/㎖)을 90분 동안 처리하였다. 전체 RNA는 트라이졸을 사용하여 추출하였고, M-MuLV 역전사 중합효소(Thermo scientific)를 사용하여 cDNA를 합성하였다. 플루코나졸 처리 또는 미처리된 세포 내 전체 RNA에 대하여 ERG 유전자 특이적인 프라이머를 사용하여 증폭시킨 각각의 ERG 유전자 특이적 탐침을 이용하여 노던 블롯 분석을 실시하였다: 프라이머; ERG2에 대한 B5789(5'-CAAGAAATGGAGCGTGAG-3') 및 B5790(5'-CAGTGTTGTAAAGCGTGATG-3'); ERG3에 대한 B1720(5'-ATCCCTTTTCACCGTCGCTC-3') 및 B1721(5'-CGTCGTGGATGAGAATAGTCC-3'); ERG5에 대한 B671(5'-GTTTTGTTGCCTGAGAACTGGG-3') 및 B674(5'-ATCACTCAACTCGGTCCTCTCGTG-3'); ERG11에 대한 B678(5'-TTCAGGGAACTTGGGAACAGC-3') 및 B1598(5'-CAGGAGCAGAAACAAAGC-3'); ERG25에 대한 B1718(5'-TGACCGCCTGTAGATTGTC-3') 및 B1719(5'-TAGTCCCACCACCTGAAAC-3'). MyiQ2 리얼-타임 PCR 탐지 시스템(Bio Rad) 및 각각의 유전자-특이적인 프라이머를 사용하여 정량적 리얼-타임 PCR을 실시하였다. 프라이머: ERG2에 대한 B5789(5'-CAAGAAATGGAGCGTGAG-3') 및 B6838(5'-GGGAGGGTCGAGGATGTAGA-3'); ERG3에 대한 B1720(5'-ATCCCTTTTCACCGTCGCTC-3') 및 B6838(5'-GGGAGGGTCGAGGATGTAGA-3'); ERG5에 대한 B671(5'-GTTTTGTTGCCTGAGAACTGGG-3') 및 B672(5'-GTAGATACTGAGAGCCTGCTTGGTG-3'); ERG11에 대한 B677(5'-AATCTCCTTACCAGCCATTCGG-3') 및 B678(5'-TTCAGGGAACTTGGGAACAGC-3'); ERG25에 대한 B2695(5'-TCGTCTTTGGCAAGCAGTC-3') 및 B6840(5'-GAAGTCGTGGTGGTCAGCA-3'); ACT1에 대한 B679(5'-CGCCCTTGCTCCTTCTTCTATG-3') 및 B680(5'-TACTCGTCGTATTCGCTCTTCG-3'). 예측된 바와 같이, ERG11의 기본 수준 및 유도 수준은 야생형 균주보다 sre1 변이체에서 매우 낮았다. 놀랍게도, HOB1 결실은 ERG11의 기본 발현 수준을 크게 증가시켰다. 스테롤-충분 조건 및 스테롤-결핍 조건에서 야생형 균주, hob1Δ 균주 및 sre1Δ 균주 내 ERG2, ERG3, ERG5 및 ERG25의 발현을 관찰하여 Hob1이 다른 ERG 유전자 조절에 관련되어 있는지를 검사하였다. 플루코나졸 처리를 통한 스테롤 결핍에 대한 반응에서 다른 모든 ERG 유전자의 발현은 야생형 균주에 유도되었으나 sre1Δ 균주에서는 유도되지 않았다(도 14c). HOB1의 결실은 ERG2의 기본 발현을 크게 증가시켰다(도 14c). 반대로, 스테롤 결핍 시에, hob1Δ 변이체 내 ERG2, ERG3, ERG5, ERG11 및 ERG25의 유도는 감소되었다(도 14c). ERG 발현의 견고한 조절은 hob1Δ 변이체 내 대부분 결핍되어 있는데, 이는 Hob1이 에르고스테롤(ergosterol) 유전자 발현의 주요 조절인자임을 암시하였다(도 14d, 도 14e). 스테롤 생합성에 관련된 전사인자는 환경 스트레스 반응 및 적응에 관련된 것으로 예측되는데, 병원균이 감염 중에 급격한 환경 변화에 직면하기 때문에 환경 스트레스에 대한 반응 및 적응은 크립토코커스 네오포만스의 생존 및 증식에서 매우 중요하다(비특허문헌 3).In the present invention, it was confirmed that the deletion of some transcription factors negatively regulates the sensitivity to azole and polyene drugs (Figs. 13 and 14A), which means that the transcription factors directly regulate ERG11 expression and sterol biosynthesis to bind polyene. Because it affects. Two of the above transcription factors are already known as Erg11 modulators. Sre1, as a major regulator of sterol, formed a complex with Scp1, which is a part of the sterol regulator-binding protein (SREBP) in Cryptococcus neoformans (
외부 스트레스 반응성 검사External stress responsiveness test
외부 스트레스에 관련된 표현형을 분석하기 위해서, 세포는 액체 YPD 배지 내 30℃에서 16시간 동안 배양하였고, 10배로 순차적 희석하였으며(1 내지 104 희석), 하기의 스트레스 유발 인자의 지정된 농도를 포함하는 YPD 배지 상에 스팟팅하였다(spotted); A: 삼투압 스트레스(소르비톨); B: 산화 스트레스[과산화수소(H2O2),터트-부틸 하이드로퍼록사이드(TH, tert-butyl hydroperoxide, 유기 퍼록사이드), 메나디온(menadione), 다이아마이드(diamide)]; C: 소포체(ER) 스트레스[튜니카마이신(tumicamycin), 디티오트레이톨(DTT, dithiothreitol)]; D: 유전독성 스트레스[메틸메탄 설페이트(MMS, methyl methanesulfonate), 하이드록시유레아(HU, hydroxyurea)]; E: 세포막/세포벽 불안정화 스트레스[칼코플루오르 화이트(CFW, calcofluor white), 콩고 레드(CR, Congo red), 소디움 도데실 설페이트(SDS, Sodium Dodecyl Sulfate)]; F: 중금속 스트레스(CdSO4). 세포는 30℃에서 배양하였고 2일 내지 5일간 촬영하였다.In order to analyze the phenotype related to external stress, cells were incubated for 16 hours at 30° C. in liquid YPD medium, and serially diluted 10-fold (1 to 10 4 dilutions), and YPD containing the specified concentration of the following stress inducing factors Spotted on the medium; A: osmotic stress (sorbitol); B: Oxidative stress [hydrogen peroxide (H 2 O 2 ), tert-butyl hydroperoxide (TH, tert-butyl hydroperoxide, organic peroxide), menadione, diamide]; C: endoplasmic reticulum (ER) stress (tumicamycin, dithiothreitol (DTT)); D: genotoxic stress [methyl methanesulfonate (MMS), hydroxyurea (HU, hydroxyurea)]; E: Cell membrane/cell wall destabilization stress (Calcofluor white (CFW), Congo red (CR), Sodium Dodecyl Sulfate (SDS)); F: heavy metal stress (CdSO 4 ). Cells were cultured at 30° C. and photographed for 2 to 5 days.
다양한 형태의 외부 스트레스에 대한 반응으로, 145가지 전사인자는 최소한 한 가지 형태의 스트레스를 인지하여 대응하는 것으로 확인되었다(도 4 및 도 15). 이러한 전사인자 중에서, 2가지 스테롤 조절인자 Sre1 및 Hob1은 복수의 스트레스 반응 및 적응을 조절하는 일반적인 스트레스-반응성 전사인자인 것으로 예측되었다. 놀랍게도, HOB1 또는 SRE1의 결실은 삼투압/염, 산화/환원, 유전독성, ER, 및 세포 벽/막 스트레스에 대한 내성을 크게 감소시켰다(도 14d). hob1 및 sre1 변이체는 대부분의 다양한 환경 스트레스에 대하여 유사한 내성 및 감수성 패턴을 나타내었는데, 이는 플루코나졸(FCZ) 및 암포테리신 B(AmpB)에 대한 반응과는 정 반대였다. 이러한 결과는 스테롤 항상성이 크립토코커스 네오포만스 내 스트레스 반응 및 적응의 조절에서 매우 중요한 요소임을 제시하였다(도 14e).In response to various types of external stress, it was confirmed that 145 transcription factors responded by recognizing at least one type of stress (FIGS. 4 and 15). Among these transcription factors, two sterol regulators Sre1 and Hob1 were predicted to be general stress-responsive transcription factors that regulate multiple stress responses and adaptations. Surprisingly, deletion of HOB1 or SRE1 significantly reduced osmotic pressure/salt, oxidation/reduction, genotoxicity, ER, and resistance to cell wall/membrane stress (FIG. 14D ). The hob1 and sre1 variants showed similar patterns of resistance and susceptibility to most of the various environmental stresses, which was the opposite of the response to fluconazole (FCZ) and amphotericin B (AmpB). These results suggested that sterol homeostasis is a very important factor in the regulation of stress response and adaptation in Cryptococcus neoformans (FIG. 14E).
기능성 상관관계 분석Functional correlation analysis
본 발명에서 2가지 가능한 생체 외 및 생체 내 표현형 조합 사이의 모든 피어슨 상관계수(PCCs, Pearson Correlation Coefficients)를 계산하여 1차종속(linear dependence) 정도를 측정함으로써 표현형적 특징과 독성 간의 상관 관계를 검증하였다. 상기 상관 관계는 결합 네트워크로 나타내었다(도 16). 상관 관계 네트워크에서 크립토코커스 네오포만스의 독성이 상이한 온도에서의 성장, 삼투압-스트레스 반응 및 세포 벽/막-스트레스 반응성과의 높은 상관관계를 나타내었고, 산화-스트레스 반응, 유전독성 스트레스 반응 및 세포벽/세포막-스트레스 반응과는 보통수준의 관련성을 나타내었다. 특히, 이러한 스트레스 표현형들은 밀접하게 교차-연관되어 있는데(도 16), 기존에 알려진 Hog1, Pkc1/Mpk1, 및 UPR 신호전달 경로를 포함하는 여러가지 중요한 스트레스 신호전달 네트워크가 있다는 것을 제시하고 있다. 대조적으로, 교배 및 항진균제(AmpB 제외) 내성은 크립토코커스 네오포만스의 독성에는 크게 연관되지 않았다. 독성 인자들의 생산은 생체 내 독성과 연관되지 않은 것처럼 나타났는데, 이는 독성인자 생산의 증가에 의해 독성이 증가하지 않기 때문인 것으로 판단되었다. 이에 대한 근거로, 캡슐 생산의 감소는 PCC 분석(P<0.05)에 의한 독성 감소와 높은 상관관계를 나타내었다.In the present invention, the correlation between phenotypic features and toxicity is verified by calculating the degree of linear dependence by calculating all Pearson Correlation Coefficients (PCCs) between two possible combinations of in vitro and in vivo phenotypes. I did. The correlation was shown as a binding network (Figure 16). In the correlation network, the toxicity of Cryptococcus neoformans showed a high correlation with growth at different temperatures, osmotic pressure-stress response, and cell wall/membrane-stress response, and oxidation-stress response, genotoxic stress response, and cell wall /The cell membrane-stress response showed a moderate level of association. In particular, these stress phenotypes are closely cross-related (FIG. 16), suggesting that there are several important stress signaling networks including the previously known Hog1, Pkc1/Mpk1, and UPR signaling pathways. In contrast, mating and antifungal (except AmpB) resistance were not significantly associated with the toxicity of Cryptococcus neoformans. The production of toxic factors appeared not to be associated with toxicity in vivo, which was judged to be because toxicity did not increase due to the increase in toxic factor production. As a basis for this, the decrease in capsule production showed a high correlation with the decrease in toxicity by PCC analysis (P<0.05).
크립토코커스Cryptococcus 네오포만스Neoformance 전사 인자에 대한 데이터베이스 구축 Build a database for transcription factors
155가지 전사인자에 대하여 수집한 유전체 자료 및 전사체 자료는 비교 진균 유전체학 플랫폼(CFGP 2.0; 비특허문헌 37) 내 표준화된 유전제 자료 내 프로토콜을 이용하여 정리하였다. 예측되는 유전자들의 상세한 정보에 대하여, 8가지 생물정보학 프로그램(InterPro scan, Signalp 3.0, PSortII, TargetP, ChloroP, SecretomeP, predictsNLS, TMHMM2)에 의해 미리-계산된 결과를 제공하였다(비특허문헌 38, 비특허문헌 39, 비특허문헌 40, 비특허문헌 41, 비특허문헌 42, 비특허문헌 43, 비특허문헌 44 및 비특허문헌 45).Genomic data and transcript data collected for 155 transcription factors were organized using a protocol within the standardized genetic data data in the comparative fungal genomics platform (CFGP 2.0; Non-Patent Document 37). For detailed information of predicted genes, pre-calculated results were provided by 8 bioinformatics programs (InterPro scan, Signalp 3.0, PSortII, TargetP, ChloroP, SecretomeP, predictsNLS, TMHMM2) (
서울대학교 유전체 브라우저(SNUGB; 비특허문헌 46)를 크립토코커스(Cryptococcus) 전사인자 데이터베이스에 병합하여, 중요한 생물학적 특징으로 유전체 내용을 검색하였다. 브라우저 스캐폴드, 유전자 모델 탐색 및 3 유전자-군 브라우저 페이지 내에서, SNUGB 모듈에 대한 직접 연결을 제공하였다. MySQL 5.0.81(소스 코드 분산) 및 PHP 5.2.6은 데이터베이스 조작 및 웹 인터페이스 개발에 각각 사용하였다. 웹 페이지는 Apache 2.2.9 웹 서버를 통해 제공하였다.By merging the Seoul National University genome browser (SNUGB; Non-Patent Document 46) into the Cryptococcus transcription factor database, genome content was searched as an important biological feature. A direct link to the SNUGB module was provided within the browser scaffold, genetic model search, and 3 gene-group browser pages. MySQL 5.0.81 (source code distribution) and PHP 5.2.6 were used for database manipulation and web interface development, respectively. The web page was provided through the Apache 2.2.9 web server.
피어슨 상관계수 네트워크 구축Pearson correlation coefficient network construction
본 발명에서 표현형 검사 결과(강한 내성 표현형:3 ; 중간 내성 표현형:2 ; 약한 내성 표현형:1 ; 야생형-유사 표현형:0 ; 약한 감수성 표현형:-1 ; 중간 감수성 표현형:-2 ; 강한 감수성 표현형:-3)을 기초로 하고 Prism 5.0(GraphPad Software, Inc)을 이용하여 피어슨 상관계수(PCC) 수치를 계산하였다. 네트워크는 PCC 수치에 기반한 Cytoscape 소프트웨어 3.2.0을 사용하여 시각화하였다.Phenotypic test results in the present invention (strongly resistant phenotype:3; moderately resistant phenotype:2; weakly resistant phenotype:1; wild type-like phenotype:0; weakly sensitive phenotype:-1; moderately sensitive phenotype:-2; strong sensitive phenotype: Pearson correlation coefficient (PCC) values were calculated based on -3) and using Prism 5.0 (GraphPad Software, Inc). The network was visualized using Cytoscape software 3.2.0 based on PCC figures.
<110> AmtixBio Co., Ltd. <120> Novel genes for regulating the virulence of Cryptococcus neoformans and their use <130> AMTIX16P-0001.2-KR <160> 1380 <170> KopatentIn 2.0 <210> 1 <211> 19 <212> DNA <213> CNAG_03132 5 flanking region primer 1 <400> 1 gcgtggcatt tagagagtg 19 <210> 2 <211> 38 <212> DNA <213> CNAG_03132 5 flanking region primer 2 <400> 2 tcactggccg tcgttttacg agcgtattgg atgaggag 38 <210> 3 <211> 39 <212> DNA <213> CNAG_03132 3 flanking region primer 1 <400> 3 catggtcata gctgtttcct gaaacctcgc tttgagacg 39 <210> 4 <211> 18 <212> DNA <213> CNAG_03132 3 flanking region primer 2 <400> 4 tgacgaagcg acagaatc 18 <210> 5 <211> 20 <212> DNA <213> CNAG_03132 diagnostic screening primer, pairing with B79 <400> 5 cccaaacact attatcgcag 20 <210> 6 <211> 18 <212> DNA <213> CNAG_03132 Southern blot probe primer <400> 6 tactcgtatg aatcggcg 18 <210> 7 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 7 tgctagaggg cgggagagtt 20 <210> 8 <211> 24 <212> DNA <213> STM common primer <400> 8 gcatgccctg cccctaagaa ttcg 24 <210> 9 <211> 20 <212> DNA <213> CNAG_05380 5 flanking region primer 1 <400> 9 cctactgctt ggcacattac 20 <210> 10 <211> 35 <212> DNA <213> CNAG_05380 5 flanking region primer 2 <400> 10 ctggccgtcg ttttaccgtc gtttcttttt gggag 35 <210> 11 <211> 37 <212> DNA <213> CNAG_05380 3 flanking region primer 1 <400> 11 gtcatagctg tttcctgtcc atctccagtc tgtgaac 37 <210> 12 <211> 20 <212> DNA <213> CNAG_05380 3 flanking region primer 2 <400> 12 cgcttcttag gacctcactc 20 <210> 13 <211> 20 <212> DNA <213> CNAG_05380 diagnostic screening primer, pairing with B79 <400> 13 tctgaaggcg gaatctggac 20 <210> 14 <211> 24 <212> DNA <213> CNAG_05380 Southern blot probe primer <400> 14 ggagcaagga actaaagatg gtag 24 <210> 15 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 15 tgctagaggg cgggagagtt 20 <210> 16 <211> 24 <212> DNA <213> STM common primer <400> 16 gcatgccctg cccctaagaa ttcg 24 <210> 17 <211> 20 <212> DNA <213> CNAG_06719 5 flanking region primer 1 <400> 17 tcgccaactg aacaaacccc 20 <210> 18 <211> 36 <212> DNA <213> CNAG_06719 5 flanking region primer 2 <400> 18 ctggccgtcg ttttactccc gcctaacact ttcgtg 36 <210> 19 <211> 36 <212> DNA <213> CNAG_06719 3 flanking region primer 1 <400> 19 gtcatagctg tttcctgtgt caagccataa agagcc 36 <210> 20 <211> 20 <212> DNA <213> CNAG_06719 3 flanking region primer 2 <400> 20 ctgatagttt tgccacttgc 20 <210> 21 <211> 22 <212> DNA <213> CNAG_06719 diagnostic screening primer, pairing with B79 <400> 21 aataccctcc ttacttcccc gc 22 <210> 22 <211> 18 <212> DNA <213> CNAG_06719 Southern blot probe primer <400> 22 gtaaaagttt gggcgtgg 18 <210> 23 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 23 tgctagaggg cgggagagtt 20 <210> 24 <211> 24 <212> DNA <213> STM common primer <400> 24 gcatgccctg cccctaagaa ttcg 24 <210> 25 <211> 20 <212> DNA <213> CNAG_04093 5 flanking region primer 1 <400> 25 tccaggcgta ttcatcccag 20 <210> 26 <211> 38 <212> DNA <213> CNAG_04093 5 flanking region primer 2 <400> 26 ctggccgtcg ttttacaagg attgtgacct caggctcg 38 <210> 27 <211> 36 <212> DNA <213> CNAG_04093 3 flanking region primer 1 <400> 27 gtcatagctg tttcctgatg tttctcctcc agagcc 36 <210> 28 <211> 19 <212> DNA <213> CNAG_04093 3 flanking region primer 2 <400> 28 gcaagaacca aatgacagc 19 <210> 29 <211> 24 <212> DNA <213> CNAG_04093 diagnostic screening primer, pairing with B79 <400> 29 atcattcctc tgacaagtcg ggcg 24 <210> 30 <211> 19 <212> DNA <213> CNAG_04093 Southern blot probe primer <400> 30 atgtgtcctg gatggcttg 19 <210> 31 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 31 tgctagaggg cgggagagtt 20 <210> 32 <211> 24 <212> DNA <213> STM common primer <400> 32 gcatgccctg cccctaagaa ttcg 24 <210> 33 <211> 18 <212> DNA <213> CNAG_04518 5 flanking region primer 1 <400> 33 tgtcctttct tgtgaccg 18 <210> 34 <211> 39 <212> DNA <213> CNAG_04518 5 flanking region primer 2 <400> 34 tcactggccg tcgttttacc agtaacgaag aacgcaatc 39 <210> 35 <211> 40 <212> DNA <213> CNAG_04518 3 flanking region primer 1 <400> 35 catggtcata gctgtttcct gcacaagcca cgattattgc 40 <210> 36 <211> 20 <212> DNA <213> CNAG_04518 3 flanking region primer 2 <400> 36 tgtatctgtg tgtggactgc 20 <210> 37 <211> 18 <212> DNA <213> CNAG_04518 diagnostic screening primer, pairing with B79 <400> 37 acacaaaggg aggaatgc 18 <210> 38 <211> 19 <212> DNA <213> CNAG_04518 Southern blot probe primer <400> 38 catacactgt gccacgaac 19 <210> 39 <211> 20 <212> DNA <213> NAT#6 STM primer <400> 39 atagctacca cacgatagct 20 <210> 40 <211> 24 <212> DNA <213> STM common primer <400> 40 gcatgccctg cccctaagaa ttcg 24 <210> 41 <211> 20 <212> DNA <213> CNAG_01858 5 flanking region primer 1 <400> 41 tcttcaggaa cataggatgg 20 <210> 42 <211> 38 <212> DNA <213> CNAG_01858 5 flanking region primer 2 <400> 42 tcactggccg tcgttttacg agattgttgg gaggattg 38 <210> 43 <211> 39 <212> DNA <213> CNAG_01858 3 flanking region primer 1 <400> 43 catggtcata gctgtttcct ggggcgaata aggtttctg 39 <210> 44 <211> 18 <212> DNA <213> CNAG_01858 3 flanking region primer 2 <400> 44 tgttttcgct catccgtc 18 <210> 45 <211> 19 <212> DNA <213> CNAG_01858 diagnostic screening primer, pairing with B79 <400> 45 aacacagcgg aatgacctc 19 <210> 46 <211> 18 <212> DNA <213> CNAG_01858 Southern blot probe primer <400> 46 cacttggttc attgctgg 18 <210> 47 <211> 20 <212> DNA <213> NAT#43 STM primer <400> 47 ccagctacca atcacgctac 20 <210> 48 <211> 24 <212> DNA <213> STM common primer <400> 48 gcatgccctg cccctaagaa ttcg 24 <210> 49 <211> 22 <212> DNA <213> CNAG_02305 5 flanking region primer 1 <400> 49 ccagaaggat gtgatactga gg 22 <210> 50 <211> 36 <212> DNA <213> CNAG_02305 5 flanking region primer 2 <400> 50 ctggccgtcg ttttacgaaa tcgttctgaa actgcg 36 <210> 51 <211> 39 <212> DNA <213> CNAG_02305 3 flanking region primer 1 <400> 51 gtcatagctg tttcctgatc gttcacaggc tcaaggctg 39 <210> 52 <211> 22 <212> DNA <213> CNAG_02305 3 flanking region primer 2 <400> 52 gaggtcatta gagggtattt gc 22 <210> 53 <211> 19 <212> DNA <213> CNAG_02305 diagnostic screening primer, pairing with B79 <400> 53 gcgtggcgaa tacatacag 19 <210> 54 <211> 19 <212> DNA <213> CNAG_02305 Southern blot probe primer <400> 54 aggtgctaca aagactcgg 19 <210> 55 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 55 cgcaaaatca ctagccctat agcg 24 <210> 56 <211> 24 <212> DNA <213> STM common primer <400> 56 gcatgccctg cccctaagaa ttcg 24 <210> 57 <211> 19 <212> DNA <213> CNAG_06156 5 flanking region primer 1 <400> 57 aagatagcac ctccgacag 19 <210> 58 <211> 41 <212> DNA <213> CNAG_06156 5 flanking region primer 2 <400> 58 gctcactggc cgtcgtttta cgctgttgcg aaagataaag g 41 <210> 59 <211> 41 <212> DNA <213> CNAG_06156 3 flanking region primer 1 <400> 59 catggtcata gctgtttcct gccaggaatg aactaccaag g 41 <210> 60 <211> 19 <212> DNA <213> CNAG_06156 3 flanking region primer 2 <400> 60 ttgaagcagg actctccac 19 <210> 61 <211> 19 <212> DNA <213> CNAG_06156 diagnostic screening primer, pairing with B79 <400> 61 tgcttgtcgt ttggttagg 19 <210> 62 <211> 19 <212> DNA <213> CNAG_06156 Southern blot probe primer <400> 62 ggtgaaaatg acggtccag 19 <210> 63 <211> 24 <212> DNA <213> NAT#119 STM primer <400> 63 ctccccacat aaagagagct aaac 24 <210> 64 <211> 24 <212> DNA <213> STM common primer <400> 64 gcatgccctg cccctaagaa ttcg 24 <210> 65 <211> 21 <212> DNA <213> CNAG_06188 5 flanking region primer 1 <400> 65 ctcttgtctt ctgcctattc c 21 <210> 66 <211> 35 <212> DNA <213> CNAG_06188 5 flanking region primer 2 <400> 66 ctggccgtcg ttttacggac atagtgaaac tgggg 35 <210> 67 <211> 38 <212> DNA <213> CNAG_06188 3 flanking region primer 1 <400> 67 gtcatagctg tttcctgtca tcctccgact ctaaactg 38 <210> 68 <211> 20 <212> DNA <213> CNAG_06188 3 flanking region primer 2 <400> 68 gttttcctac ttttgtgggc 20 <210> 69 <211> 20 <212> DNA <213> CNAG_06188 diagnostic screening primer, pairing with B79 <400> 69 agaagacgac tgtggtaggc 20 <210> 70 <211> 20 <212> DNA <213> CNAG_06188 Southern blot probe primer <400> 70 gagtcagcag cagaagattc 20 <210> 71 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 71 acagctccaa acctcgctaa acag 24 <210> 72 <211> 24 <212> DNA <213> STM common primer <400> 72 gcatgccctg cccctaagaa ttcg 24 <210> 73 <211> 19 <212> DNA <213> CNAG_05785 5 flanking region primer 1 <400> 73 catttctact tggttgccg 19 <210> 74 <211> 36 <212> DNA <213> CNAG_05785 5 flanking region primer 2 <400> 74 ctggccgtcg ttttacagga gaaagtgatg gtcggc 36 <210> 75 <211> 39 <212> DNA <213> CNAG_05785 3 flanking region primer 1 <400> 75 gtcatagctg tttcctgtta tcgtgagcct tcaagcccc 39 <210> 76 <211> 21 <212> DNA <213> CNAG_05785 3 flanking region primer 2 <400> 76 ccattgtaga tgtcggttac g 21 <210> 77 <211> 22 <212> DNA <213> CNAG_05785 diagnostic screening primer, pairing with B79 <400> 77 ttgcggtctt tgtcccttgt gg 22 <210> 78 <211> 24 <212> DNA <213> CNAG_05785 Southern blot probe primer <400> 78 ttctccactt gctcgcctat cctc 24 <210> 79 <211> 24 <212> DNA <213> NAT#125 STM primer <400> 79 cgctacagcc agcgcgcgca agcg 24 <210> 80 <211> 24 <212> DNA <213> STM common primer <400> 80 gcatgccctg cccctaagaa ttcg 24 <210> 81 <211> 20 <212> DNA <213> CNAG_05019 5 flanking region primer 1 <400> 81 aaccaccaca atgtcttcag 20 <210> 82 <211> 40 <212> DNA <213> CNAG_05019 5 flanking region primer 2 <400> 82 gctcactggc cgtcgtttta cagacccttg tttggttctg 40 <210> 83 <211> 41 <212> DNA <213> CNAG_05019 3 flanking region primer 1 <400> 83 catggtcata gctgtttcct gagtggagag ggagagcaat g 41 <210> 84 <211> 18 <212> DNA <213> CNAG_05019 3 flanking region primer 2 <400> 84 tttacgaaga gcagcgac 18 <210> 85 <211> 18 <212> DNA <213> CNAG_05019 diagnostic screening primer, pairing with B79 <400> 85 agatagcaac ggctttgg 18 <210> 86 <211> 20 <212> DNA <213> CNAG_05019 Southern blot probe primer <400> 86 cgcaaaaggg aagatggata 20 <210> 87 <211> 20 <212> DNA <213> NAT#150 STM primer <400> 87 acatacaccc ccatcccccc 20 <210> 88 <211> 24 <212> DNA <213> STM common primer <400> 88 gcatgccctg cccctaagaa ttcg 24 <210> 89 <211> 18 <212> DNA <213> CNAG_02877 5 flanking region primer 1 <400> 89 gtttaccatt tcaccgcc 18 <210> 90 <211> 39 <212> DNA <213> CNAG_02877 5 flanking region primer 2 <400> 90 tcactggccg tcgttttact gatgaggata cattcggag 39 <210> 91 <211> 41 <212> DNA <213> CNAG_02877 3 flanking region primer 1 <400> 91 catggtcata gctgtttcct gggcttcaac aatcaccatt c 41 <210> 92 <211> 18 <212> DNA <213> CNAG_02877 3 flanking region primer 2 <400> 92 acgaaaagcg gatagcag 18 <210> 93 <211> 20 <212> DNA <213> CNAG_02877 diagnostic screening primer, pairing with B79 <400> 93 agccttatta tgaccaaccc 20 <210> 94 <211> 20 <212> DNA <213> CNAG_02877 Southern blot probe primer <400> 94 caagttcctt tgcctacacc 20 <210> 95 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 95 acgcaccaga cacacaacca g 21 <210> 96 <211> 24 <212> DNA <213> STM common primer <400> 96 gcatgccctg cccctaagaa ttcg 24 <210> 97 <211> 20 <212> DNA <213> CNAG_00039 5 flanking region primer 1 <400> 97 tcactctttc ggggcatcag 20 <210> 98 <211> 38 <212> DNA <213> CNAG_00039 5 flanking region primer 2 <400> 98 ctggccgtcg ttttacctaa ctggggatga aatgtctg 38 <210> 99 <211> 39 <212> DNA <213> CNAG_00039 3 flanking region primer 1 <400> 99 gtcatagctg tttcctgggg gaagagcaga aagagtaac 39 <210> 100 <211> 22 <212> DNA <213> CNAG_00039 3 flanking region primer 2 <400> 100 gcaatcgtaa acaacaccct tg 22 <210> 101 <211> 20 <212> DNA <213> CNAG_00039 diagnostic screening primer, pairing with B79 <400> 101 cgtctgtcgt ttatcggttg 20 <210> 102 <211> 19 <212> DNA <213> CNAG_00039 Southern blot probe primer <400> 102 accctgtaac atttgtccg 19 <210> 103 <211> 19 <212> DNA <213> CNAG_00039 Southern blot probe primer <400> 103 acttaccctt cacgccaag 19 <210> 104 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 104 caccaactcc ccatctccat 20 <210> 105 <211> 24 <212> DNA <213> STM common primer <400> 105 gcatgccctg cccctaagaa ttcg 24 <210> 106 <211> 20 <212> DNA <213> CNAG_00039 5 flanking region primer 1 <400> 106 agacattggg cttgacggtg 20 <210> 107 <211> 39 <212> DNA <213> CNAG_00039 5 flanking region primer 2 <400> 107 tcactggccg tcgttttaca cgagttggag aggggttac 39 <210> 108 <211> 39 <212> DNA <213> CNAG_00039 3 flanking region primer 1 <400> 108 catggtcata gctgtttcct gtagaaagga atcggggac 39 <210> 109 <211> 20 <212> DNA <213> CNAG_00039 3 flanking region primer 2 <400> 109 ggagaacgaa aacaaagtgc 20 <210> 110 <211> 19 <212> DNA <213> CNAG_00039 diagnostic screening primer, pairing with B79 <400> 110 ggttcttgtt tatgccctg 19 <210> 111 <211> 20 <212> DNA <213> CNAG_00039 Southern blot probe primer <400> 111 aacgagggta gattgcgtcg 20 <210> 112 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 112 caccaactcc ccatctccat 20 <210> 113 <211> 24 <212> DNA <213> STM common primer <400> 113 gcatgccctg cccctaagaa ttcg 24 <210> 114 <211> 19 <212> DNA <213> CNAG_01014 5 flanking region primer 1 <400> 114 tatcgccgac atctctcag 19 <210> 115 <211> 38 <212> DNA <213> CNAG_01014 5 flanking region primer 2 <400> 115 tcactggccg tcgttttact ttaggacgga ggggattg 38 <210> 116 <211> 39 <212> DNA <213> CNAG_01014 3 flanking region primer 1 <400> 116 catggtcata gctgtttcct ggagccgaat caggaaatc 39 <210> 117 <211> 19 <212> DNA <213> CNAG_01014 3 flanking region primer 2 <400> 117 cgcaggcttt tctactctc 19 <210> 118 <211> 30 <212> DNA <213> CNAG_01014 diagnostic screening primer, pairing with B79 <400> 118 cgcgcggccg cgaaaacctc cacagagcag 30 <210> 119 <211> 19 <212> DNA <213> CNAG_01014 Southern blot probe primer <400> 119 ttgcttgctg tctcgtaac 19 <210> 120 <211> 18 <212> DNA <213> CNAG_01014 Southern blot probe primer <400> 120 gatttcctga ttcggctc 18 <210> 121 <211> 20 <212> DNA <213> CTAGAGCCCGCCACAACGCT <400> 121 ctagagcccg ccacaacgct 20 <210> 122 <211> 24 <212> DNA <213> STM common primer <400> 122 gcatgccctg cccctaagaa ttcg 24 <210> 123 <211> 21 <212> DNA <213> CNAG_06762 5 flanking region primer 1 <400> 123 tgggtgagag aaagatgtca g 21 <210> 124 <211> 39 <212> DNA <213> CNAG_06762 5 flanking region primer 2 <400> 124 tcactggccg tcgttttact gtgagtggtt gtgctgtgg 39 <210> 125 <211> 41 <212> DNA <213> CNAG_06762 3 flanking region primer 1 <400> 125 catggtcata gctgtttcct gggctcatag gtgttgtaac g 41 <210> 126 <211> 20 <212> DNA <213> CNAG_06762 3 flanking region primer 2 <400> 126 agtattcggt ggtgacaaag 20 <210> 127 <211> 21 <212> DNA <213> CNAG_06762 diagnostic screening primer, pairing with B79 <400> 127 tgggtgagag aaagatgtca g 21 <210> 128 <211> 19 <212> DNA <213> CNAG_06762 Southern blot probe primer <400> 128 tgttccaccc cttgctaac 19 <210> 129 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 129 ctccacatcc atcgctccaa 20 <210> 130 <211> 24 <212> DNA <213> STM common primer <400> 130 gcatgccctg cccctaagaa ttcg 24 <210> 131 <211> 18 <212> DNA <213> CNAG_04630 5 flanking region primer 1 <400> 131 atggaaggac tttgacgg 18 <210> 132 <211> 40 <212> DNA <213> CNAG_04630 5 flanking region primer 2 <400> 132 tcactggccg tcgttttact gactcacact tgattctccc 40 <210> 133 <211> 40 <212> DNA <213> CNAG_04630 3 flanking region primer 1 <400> 133 gatggtcata gctgtttcct gctcattttc cctttcgtcg 40 <210> 134 <211> 18 <212> DNA <213> CNAG_04630 3 flanking region primer 2 <400> 134 taaaggaatg cggctgag 18 <210> 135 <211> 19 <212> DNA <213> CNAG_04630 diagnostic screening primer, pairing with B79 <400> 135 ccaacagcaa cttcgtttc 19 <210> 136 <211> 21 <212> DNA <213> CNAG_04630 Southern blot probe primer <400> 136 tctcttctcc ttggaactga g 21 <210> 137 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 137 ctccacatcc atcgctccaa 20 <210> 138 <211> 24 <212> DNA <213> STM common primer <400> 138 gcatgccctg cccctaagaa ttcg 24 <210> 139 <211> 19 <212> DNA <213> CNAG_03401 5 flanking region primer 1 <400> 139 ttcctcgtcc aagtaaacc 19 <210> 140 <211> 41 <212> DNA <213> CNAG_03401 5 flanking region primer 2 <400> 140 gctcactggc cgtcgtttta caatctcgca aaccgcacac g 41 <210> 141 <211> 43 <212> DNA <213> CNAG_03401 3 flanking region primer 1 <400> 141 catggtcata gctgtttcct gagcgcgaac agggaaatac acc 43 <210> 142 <211> 18 <212> DNA <213> CNAG_03401 3 flanking region primer 2 <400> 142 tcgtgtagga atgggtag 18 <210> 143 <211> 20 <212> DNA <213> CNAG_03401 diagnostic screening primer, pairing with B79 <400> 143 tggtctcccc gacgatttac 20 <210> 144 <211> 19 <212> DNA <213> CNAG_03401 Southern blot probe primer <400> 144 tatggtcagg aggaagcag 19 <210> 145 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 145 cagatctcga acgataccca 20 <210> 146 <211> 24 <212> DNA <213> STM common primer <400> 146 gcatgccctg cccctaagaa ttcg 24 <210> 147 <211> 22 <212> DNA <213> CNAG_04594 5 flanking region primer 1 <400> 147 tggcatcgtg ctgttacaag tc 22 <210> 148 <211> 45 <212> DNA <213> CNAG_04594 5 flanking region primer 2 <400> 148 gctcactggc cgtcgtttta catttgcgga tggacctgcc ttagg 45 <210> 149 <211> 41 <212> DNA <213> CNAG_04594 3 flanking region primer 1 <400> 149 catggtcata gctgtttcct gatcaggatt caggacacga c 41 <210> 150 <211> 18 <212> DNA <213> CNAG_04594 3 flanking region primer 2 <400> 150 tttggttctc tctcccac 18 <210> 151 <211> 19 <212> DNA <213> CNAG_04594 diagnostic screening primer, pairing with B79 <400> 151 attgcttgag accagcgtc 19 <210> 152 <211> 18 <212> DNA <213> CNAG_04594 Southern blot probe primer <400> 152 acggttgctt tgctatgc 18 <210> 153 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 153 cagatctcga acgataccca 20 <210> 154 <211> 24 <212> DNA <213> STM common primer <400> 154 gcatgccctg cccctaagaa ttcg 24 <210> 155 <211> 19 <212> DNA <213> CNAG_07901 5 flanking region primer 1 <400> 155 actcaccttc ccgctttag 19 <210> 156 <211> 34 <212> DNA <213> CNAG_07901 5 flanking region primer 2 <400> 156 ctggccgtcg ttttacgttt gagttgaggt tgcg 34 <210> 157 <211> 36 <212> DNA <213> CNAG_07901 3 flanking region primer 1 <400> 157 gtcatagctg tttcctgaga atgttgatgg tagcgg 36 <210> 158 <211> 19 <212> DNA <213> CNAG_07901 3 flanking region primer 2 <400> 158 tccaccgaag aaactgaag 19 <210> 159 <211> 18 <212> DNA <213> CNAG_07901 diagnostic screening primer, pairing with B79 <400> 159 gggatttgga tacgcttg 18 <210> 160 <211> 18 <212> DNA <213> CNAG_07901 Southern blot probe primer <400> 160 ttgcggtggg aaagtatc 18 <210> 161 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 161 ccatagaact agctaaagca 20 <210> 162 <211> 24 <212> DNA <213> STM common primer <400> 162 gcatgccctg cccctaagaa ttcg 24 <210> 163 <211> 19 <212> DNA <213> CNAG_04588 5 flanking region primer 1 <400> 163 aaaagaggtc cgatgagtg 19 <210> 164 <211> 40 <212> DNA <213> CNAG_04588 5 flanking region primer 2 <400> 164 gctcactggc cgtcgtttta cgtgaatgat aaaggcgacc 40 <210> 165 <211> 39 <212> DNA <213> CNAG_04588 3 flanking region primer 1 <400> 165 catggtcata gctgtttcct gtgctgtttc actatccgc 39 <210> 166 <211> 19 <212> DNA <213> CNAG_04588 3 flanking region primer 2 <400> 166 cgaaagcgac tggagtaac 19 <210> 167 <211> 20 <212> DNA <213> CNAG_04588 diagnostic screening primer, pairing with B79 <400> 167 gaagatgctg aagcgtattc 20 <210> 168 <211> 18 <212> DNA <213> CNAG_04588 Southern blot probe primer <400> 168 acacttatcc gccattcc 18 <210> 169 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 169 ccatagaact agctaaagca 20 <210> 170 <211> 24 <212> DNA <213> STM common primer <400> 170 gcatgccctg cccctaagaa ttcg 24 <210> 171 <211> 18 <212> DNA <213> CNAG_03998 5 flanking region primer 1 <400> 171 gccattgtga aagcatag 18 <210> 172 <211> 43 <212> DNA <213> CNAG_03998 5 flanking region primer 2 <400> 172 gctcactggc cgtcgtttta cacctgtcta tctcactgtc cag 43 <210> 173 <211> 39 <212> DNA <213> CNAG_03998 3 flanking region primer 1 <400> 173 catggtcata gctgtttcct gtgttcagca gcaaaaggc 39 <210> 174 <211> 18 <212> DNA <213> CNAG_03998 3 flanking region primer 2 <400> 174 gcggagagaa ggattatc 18 <210> 175 <211> 20 <212> DNA <213> CNAG_03998 diagnostic screening primer, pairing with B79 <400> 175 gctgacaact tggaatcatc 20 <210> 176 <211> 20 <212> DNA <213> CNAG_03998 Southern blot probe primer <400> 176 gggcttctct tcttcttcac 20 <210> 177 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 177 ccagccagca ctccgatata 20 <210> 178 <211> 24 <212> DNA <213> STM common primer <400> 178 gcatgccctg cccctaagaa ttcg 24 <210> 179 <211> 20 <212> DNA <213> CNAG_04804 5 flanking region primer 1 <400> 179 taggtagttc ctgctcgtgg 20 <210> 180 <211> 40 <212> DNA <213> CNAG_04804 5 flanking region primer 2 <400> 180 tcactggccg tcgttttacc atcttgtcct gtaatgaggc 40 <210> 181 <211> 39 <212> DNA <213> CNAG_04804 3 flanking region primer 1 <400> 181 catggtcata gctgtttcct ggattggagg gcgatttag 39 <210> 182 <211> 20 <212> DNA <213> CNAG_04804 3 flanking region primer 2 <400> 182 caacaaggtc caagaaactg 20 <210> 183 <211> 21 <212> DNA <213> CNAG_04804 diagnostic screening primer, pairing with B79 <400> 183 agaggactga gacttttgtc g 21 <210> 184 <211> 19 <212> DNA <213> CNAG_04804 Southern blot probe primer <400> 184 cgaaggggaa ctggttatc 19 <210> 185 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 185 ggtgttggat cggggtggat 20 <210> 186 <211> 24 <212> DNA <213> STM common primer <400> 186 gcatgccctg cccctaagaa ttcg 24 <210> 187 <211> 20 <212> DNA <213> CNAG_06134 5 flanking region primer 1 <400> 187 gtttgaggct ggtaaaaagg 20 <210> 188 <211> 39 <212> DNA <213> CNAG_06134 5 flanking region primer 2 <400> 188 gctcactggc cgtcgtttta catggggaat gaaagcgtg 39 <210> 189 <211> 41 <212> DNA <213> CNAG_06134 3 flanking region primer 1 <400> 189 catggtcata gctgtttcct gaaggggcga gagtagttca g 41 <210> 190 <211> 21 <212> DNA <213> CNAG_06134 3 flanking region primer 2 <400> 190 gactgtaaag gagggcataa g 21 <210> 191 <211> 19 <212> DNA <213> CNAG_06134 diagnostic screening primer, pairing with B79 <400> 191 aactcttctc agccttcgg 19 <210> 192 <211> 19 <212> DNA <213> CNAG_06134 Southern blot probe primer <400> 192 cgttctccgt cttgatagc 19 <210> 193 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 193 acacctacat caaaccctcc c 21 <210> 194 <211> 24 <212> DNA <213> STM common primer <400> 194 gcatgccctg cccctaagaa ttcg 24 <210> 195 <211> 19 <212> DNA <213> CNAG_04798 5 flanking region primer 1 <400> 195 caatgattag gcttgcgtc 19 <210> 196 <211> 38 <212> DNA <213> CNAG_04798 5 flanking region primer 2 <400> 196 tcactggccg tcgttttact gttgttgggc agtggtag 38 <210> 197 <211> 39 <212> DNA <213> CNAG_04798 3 flanking region primer 1 <400> 197 catggtcata gctgtttcct gtttagggaa gaggaagcg 39 <210> 198 <211> 18 <212> DNA <213> CNAG_04798 3 flanking region primer 2 <400> 198 tgtcatccag cagattgg 18 <210> 199 <211> 19 <212> DNA <213> CNAG_04798 diagnostic screening primer, pairing with B79 <400> 199 tgaatgactg atggacccc 19 <210> 200 <211> 18 <212> DNA <213> CNAG_04798 Southern blot probe primer <400> 200 tcggttctcg ttttggag 18 <210> 201 <211> 20 <212> DNA <213> CNAG_04864 5 flanking region primer 1 <400> 201 tcatttcttc gtcctctgag 20 <210> 202 <211> 36 <212> DNA <213> CNAG_04864 5 flanking region primer 2 <400> 202 ctggccgtcg ttttacgctc cccataatca gactcc 36 <210> 203 <211> 36 <212> DNA <213> CNAG_04864 3 flanking region primer 1 <400> 203 gtcatagctg tttcctgacg aatggagttt ctggag 36 <210> 204 <211> 20 <212> DNA <213> CNAG_04864 3 flanking region primer 2 <400> 204 gcgttctcca tagttgtagc 20 <210> 205 <211> 20 <212> DNA <213> CNAG_04864 diagnostic screening primer, pairing with B79 <400> 205 atgtagataa agcacgcacc 20 <210> 206 <211> 20 <212> DNA <213> CNAG_04864 Southern blot probe primer <400> 206 cgagttattg taggttgggc 20 <210> 207 <211> 20 <212> DNA <213> CNAG_01973 5 flanking region primer 1 <400> 207 gcttgaggtt gaaacaggtc 20 <210> 208 <211> 41 <212> DNA <213> CNAG_01973 5 flanking region primer 2 <400> 208 tcactggccg tcgttttacc agatgtgtgt aagagggtca g 41 <210> 209 <211> 39 <212> DNA <213> CNAG_01973 3 flanking region primer 1 <400> 209 catggtcata gctgtttcct gtaccgagaa cgatttggc 39 <210> 210 <211> 20 <212> DNA <213> CNAG_01973 3 flanking region primer 2 <400> 210 tcccataaga ccacacactg 20 <210> 211 <211> 19 <212> DNA <213> CNAG_01973 diagnostic screening primer, pairing with B79 <400> 211 tgatgctgaa tagacgggg 19 <210> 212 <211> 19 <212> DNA <213> CNAG_01973 Southern blot probe primer <400> 212 agccttgacg acatacctg 19 <210> 213 <211> 20 <212> DNA <213> NAT#6 STM primer <400> 213 atagctacca cacgatagct 20 <210> 214 <211> 24 <212> DNA <213> STM common primer <400> 214 gcatgccctg cccctaagaa ttcg 24 <210> 215 <211> 19 <212> DNA <213> CNAG_03914 5 flanking region primer 1 <400> 215 gatgtcgttg ttccagagc 19 <210> 216 <211> 35 <212> DNA <213> CNAG_03914 5 flanking region primer 2 <400> 216 ctggccgtcg ttttactttg gatgtaagag gtcgg 35 <210> 217 <211> 37 <212> DNA <213> CNAG_03914 3 flanking region primer 1 <400> 217 gtcatagctg tttcctgcga gtatccttac cgtgtcg 37 <210> 218 <211> 19 <212> DNA <213> CNAG_03914 3 flanking region primer 2 <400> 218 acactcctgc ccaagaaac 19 <210> 219 <211> 19 <212> DNA <213> CNAG_03914 diagnostic screening primer, pairing with B79 <400> 219 actccactcc atccaaagc 19 <210> 220 <211> 20 <212> DNA <213> CNAG_03914 Southern blot probe primer <400> 220 ataagaggtc cttttgctcc 20 <210> 221 <211> 20 <212> DNA <213> NAT#43 STM primer <400> 221 ccagctacca atcacgctac 20 <210> 222 <211> 24 <212> DNA <213> STM common primer <400> 222 gcatgccctg cccctaagaa ttcg 24 <210> 223 <211> 19 <212> DNA <213> CNAG_00828 5 flanking region primer 1 <400> 223 accagtggca aggttagag 19 <210> 224 <211> 39 <212> DNA <213> CNAG_00828 5 flanking region primer 2 <400> 224 tcactggccg tcgttttact ggcagaagca caagattag 39 <210> 225 <211> 40 <212> DNA <213> CNAG_00828 3 flanking region primer 1 <400> 225 catggtcata gctgtttcct ggccatttct cgtagcgaac 40 <210> 226 <211> 19 <212> DNA <213> CNAG_00828 3 flanking region primer 2 <400> 226 actgcttatc cccacatcg 19 <210> 227 <211> 20 <212> DNA <213> CNAG_00828 diagnostic screening primer, pairing with B79 <400> 227 tgtgaagtgt gatgatgtcc 20 <210> 228 <211> 19 <212> DNA <213> CNAG_00828 Southern blot probe primer <400> 228 ggttgccact gataaaagc 19 <210> 229 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 229 cgcaaaatca ctagccctat agcg 24 <210> 230 <211> 24 <212> DNA <213> STM common primer <400> 230 gcatgccctg cccctaagaa ttcg 24 <210> 231 <211> 19 <212> DNA <213> CNAG_00871 5 flanking region primer 1 <400> 231 atagcgggca agtgtctac 19 <210> 232 <211> 38 <212> DNA <213> CNAG_00871 5 flanking region primer 2 <400> 232 tcactggccg tcgttttacc caacctttca ttccagtc 38 <210> 233 <211> 42 <212> DNA <213> CNAG_00871 3 flanking region primer 1 <400> 233 catggtcata gctgtttcct gggatacaaa tctcaacccc tg 42 <210> 234 <211> 19 <212> DNA <213> CNAG_00871 3 flanking region primer 2 <400> 234 ggttttcact tcaagagcg 19 <210> 235 <211> 18 <212> DNA <213> CNAG_00871 diagnostic screening primer, pairing with B79 <400> 235 tccaatagca ccacaacc 18 <210> 236 <211> 19 <212> DNA <213> CNAG_00871 Southern blot probe primer <400> 236 aacatccctt tactcggcg 19 <210> 237 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 237 ccatagcgat atctacccca atct 24 <210> 238 <211> 24 <212> DNA <213> STM common primer <400> 238 gcatgccctg cccctaagaa ttcg 24 <210> 239 <211> 19 <212> DNA <213> CNAG_07464 5 flanking region primer 1 <400> 239 ggcatcagga tagaaacgc 19 <210> 240 <211> 41 <212> DNA <213> CNAG_07464 5 flanking region primer 2 <400> 240 gctcactggc cgtcgtttta cggggtgaga tttgaaggta g 41 <210> 241 <211> 40 <212> DNA <213> CNAG_07464 3 flanking region primer 1 <400> 241 catggtcata gctgtttcct ggatgtgact tggtcttggg 40 <210> 242 <211> 20 <212> DNA <213> CNAG_07464 3 flanking region primer 2 <400> 242 tggtccagtc tcctcattac 20 <210> 243 <211> 18 <212> DNA <213> CNAG_07464 diagnostic screening primer, pairing with B79 <400> 243 tcttccatct cagcatcg 18 <210> 244 <211> 24 <212> DNA <213> CNAG_07464 Southern blot probe primer <400> 244 ccatagcgat atctacccca atct 24 <210> 245 <211> 20 <212> DNA <213> NAT#150 STM primer <400> 245 acatacaccc ccatcccccc 20 <210> 246 <211> 24 <212> DNA <213> STM common primer <400> 246 gcatgccctg cccctaagaa ttcg 24 <210> 247 <211> 23 <212> DNA <213> CNAG_00460 5 flanking region primer 1 <400> 247 cgtgggagag tgagtaaagt atg 23 <210> 248 <211> 42 <212> DNA <213> CNAG_00460 5 flanking region primer 2 <400> 248 gctcactggc cgtcgtttta ccgggtaaga gtggatgtat gg 42 <210> 249 <211> 39 <212> DNA <213> CNAG_00460 3 flanking region primer 1 <400> 249 catggtcata gctgtttcct gaagggcacc gttcattag 39 <210> 250 <211> 19 <212> DNA <213> CNAG_00460 3 flanking region primer 2 <400> 250 ttcttaggat gcgggactc 19 <210> 251 <211> 18 <212> DNA <213> CNAG_00460 diagnostic screening primer, pairing with B79 <400> 251 tgcgttgaca aagtcgtg 18 <210> 252 <211> 22 <212> DNA <213> CNAG_00460 Southern blot probe primer <400> 252 tagactccag taaggtccaa tc 22 <210> 253 <211> 24 <212> DNA <213> NAT#213 STM primer <400> 253 ctggggattt tgatgtgtct atgt 24 <210> 254 <211> 24 <212> DNA <213> STM common primer <400> 254 gcatgccctg cccctaagaa ttcg 24 <210> 255 <211> 18 <212> DNA <213> CNAG_01431 5 flanking region primer 1 <400> 255 tccatttggg ggtttcag 18 <210> 256 <211> 39 <212> DNA <213> CNAG_01431 5 flanking region primer 2 <400> 256 tcactggccg tcgttttact gtcttcgttc ttgccctcc 39 <210> 257 <211> 42 <212> DNA <213> CNAG_01431 3 flanking region primer 1 <400> 257 catggtcata gctgtttcct gcgttgacag aagaggacaa gg 42 <210> 258 <211> 20 <212> DNA <213> CNAG_01431 3 flanking region primer 2 <400> 258 tgacagatga tgagcagagg 20 <210> 259 <211> 19 <212> DNA <213> CNAG_01431 diagnostic screening primer, pairing with B79 <400> 259 attccagagc ctcacttgc 19 <210> 260 <211> 21 <212> DNA <213> CNAG_01431 Southern blot probe primer <400> 260 cctgcttgtc actacactga g 21 <210> 261 <211> 24 <212> DNA <213> NAT#213 STM primer <400> 261 ctggggattt tgatgtgtct atgt 24 <210> 262 <211> 24 <212> DNA <213> STM common primer <400> 262 gcatgccctg cccctaagaa ttcg 24 <210> 263 <211> 21 <212> DNA <213> CNAG_03894 5 flanking region primer 1 <400> 263 catctatctc aacggggtat c 21 <210> 264 <211> 39 <212> DNA <213> CNAG_03894 5 flanking region primer 2 <400> 264 gctcactggc cgtcgtttta ctttttcggg agaacgcag 39 <210> 265 <211> 40 <212> DNA <213> CNAG_03894 3 flanking region primer 1 <400> 265 catggtcata gctgtttcct gaggaccaag aatgctgagg 40 <210> 266 <211> 22 <212> DNA <213> CNAG_03894 3 flanking region primer 2 <400> 266 ttctcccttg tcgccgataa gc 22 <210> 267 <211> 20 <212> DNA <213> CNAG_03894 diagnostic screening primer, pairing with B79 <400> 267 tcttatcgct caccacatcc 20 <210> 268 <211> 19 <212> DNA <213> CNAG_03894 Southern blot probe primer <400> 268 ggaacattgg gaaaaggtg 19 <210> 269 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 269 cagatctcga acgataccca 20 <210> 270 <211> 24 <212> DNA <213> STM common primer <400> 270 gcatgccctg cccctaagaa ttcg 24 <210> 271 <211> 18 <212> DNA <213> CNAG_06483 5 flanking region primer 1 <400> 271 tggagcagat ggagtttg 18 <210> 272 <211> 35 <212> DNA <213> CNAG_06483 5 flanking region primer 2 <400> 272 ctggccgtcg ttttacgctt ctccctattt ccagc 35 <210> 273 <211> 36 <212> DNA <213> CNAG_06483 3 flanking region primer 1 <400> 273 gtcatagctg tttcctgaat ggagcccaga gaacag 36 <210> 274 <211> 19 <212> DNA <213> CNAG_06483 3 flanking region primer 2 <400> 274 tctcgctgag ggaaaactc 19 <210> 275 <211> 18 <212> DNA <213> CNAG_06483 diagnostic screening primer, pairing with B79 <400> 275 ccctttctct gcgttttg 18 <210> 276 <211> 18 <212> DNA <213> CNAG_06483 Southern blot probe primer <400> 276 caatgaagaa ctggtgcg 18 <210> 277 <211> 20 <212> DNA <213> NAT#227 STM primer <400> 277 tcgtggttta gagggagcgc 20 <210> 278 <211> 24 <212> DNA <213> STM common primer <400> 278 gcatgccctg cccctaagaa ttcg 24 <210> 279 <211> 18 <212> DNA <213> CNAG_00896 5 flanking region primer 1 <400> 279 aaagggtcac gatggaac 18 <210> 280 <211> 35 <212> DNA <213> CNAG_00896 5 flanking region primer 2 <400> 280 ctggccgtcg ttttacgcga ttctttactc ctggc 35 <210> 281 <211> 36 <212> DNA <213> CNAG_00896 3 flanking region primer 1 <400> 281 gtcatagctg tttcctgttc tgagacctcc gtaacc 36 <210> 282 <211> 19 <212> DNA <213> CNAG_00896 3 flanking region primer 2 <400> 282 ctgtatcgct ttagccacc 19 <210> 283 <211> 19 <212> DNA <213> CNAG_00896 diagnostic screening primer, pairing with B79 <400> 283 tacaaggacc tcacgaacg 19 <210> 284 <211> 20 <212> DNA <213> CNAG_00896 Southern blot probe primer <400> 284 acgagcagag ggacagatag 20 <210> 285 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 285 gagagatccc aacatcacgc 20 <210> 286 <211> 24 <212> DNA <213> STM common primer <400> 286 gcatgccctg cccctaagaa ttcg 24 <210> 287 <211> 18 <212> DNA <213> CNAG_01977 5 flanking region primer 1 <400> 287 tgggaaaacg aagcactc 18 <210> 288 <211> 35 <212> DNA <213> CNAG_01977 5 flanking region primer 2 <400> 288 ctggccgtcg ttttactagg aggtcacttt cgctc 35 <210> 289 <211> 36 <212> DNA <213> CNAG_01977 3 flanking region primer 1 <400> 289 gtcatagctg tttcctgcgg caaagaacaa ggtatg 36 <210> 290 <211> 20 <212> DNA <213> CNAG_01977 3 flanking region primer 2 <400> 290 tgatgggctt gaacgaacgc 20 <210> 291 <211> 19 <212> DNA <213> CNAG_01977 diagnostic screening primer, pairing with B79 <400> 291 tgtcggcaca catctactg 19 <210> 292 <211> 22 <212> DNA <213> CNAG_01977 Southern blot probe primer <400> 292 gtctcagcct cttgttacag at 22 <210> 293 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 293 gagagatccc aacatcacgc 20 <210> 294 <211> 24 <212> DNA <213> STM common primer <400> 294 gcatgccctg cccctaagaa ttcg 24 <210> 295 <211> 19 <212> DNA <213> CNAG_01069 5 flanking region primer 1 <400> 295 cttttatcta tggcgagcg 19 <210> 296 <211> 39 <212> DNA <213> CNAG_01069 5 flanking region primer 2 <400> 296 gctcactggc cgtcgtttta ctatcgcact gctggaatc 39 <210> 297 <211> 40 <212> DNA <213> CNAG_01069 3 flanking region primer 1 <400> 297 catggtcata gctgtttcct gctactttcc ctcacatccg 40 <210> 298 <211> 19 <212> DNA <213> CNAG_01069 3 flanking region primer 2 <400> 298 tggagggtag atgcgttac 19 <210> 299 <211> 20 <212> DNA <213> CNAG_01069 diagnostic screening primer, pairing with B79 <400> 299 agagaagcgg aagagaaaac 20 <210> 300 <211> 20 <212> DNA <213> CNAG_01069 Southern blot probe primer <400> 300 ccttgcgaaa atgtcttctc 20 <210> 301 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 301 atttcgatgt atatatgtag c 21 <210> 302 <211> 24 <212> DNA <213> STM common primer <400> 302 gcatgccctg cccctaagaa ttcg 24 <210> 303 <211> 19 <212> DNA <213> CNAG_06276 5 flanking region primer 1 <400> 303 tcaagtctac gcagtcgtg 19 <210> 304 <211> 40 <212> DNA <213> CNAG_06276 5 flanking region primer 2 <400> 304 gctcactggc cgtcgtttta cgcagcaatc aactcactcg 40 <210> 305 <211> 41 <212> DNA <213> CNAG_06276 3 flanking region primer 1 <400> 305 catggtcata gctgtttcct gaggagagtg ggagtatgga g 41 <210> 306 <211> 18 <212> DNA <213> CNAG_06276 3 flanking region primer 2 <400> 306 acccatcaaa aggcgtag 18 <210> 307 <211> 20 <212> DNA <213> CNAG_06276 diagnostic screening primer, pairing with B79 <400> 307 accatttgtc aactgtcagg 20 <210> 308 <211> 20 <212> DNA <213> CNAG_06276 Southern blot probe primer <400> 308 tcgctcactg acaagaagac 20 <210> 309 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 309 atttcgatgt atatatgtag c 21 <210> 310 <211> 24 <212> DNA <213> STM common primer <400> 310 gcatgccctg cccctaagaa ttcg 24 <210> 311 <211> 19 <212> DNA <213> CNAG_02476 5 flanking region primer 1 <400> 311 tctgctcaat gaaacgggc 19 <210> 312 <211> 45 <212> DNA <213> CNAG_02476 5 flanking region primer 2 <400> 312 gctcactggc cgtcgtttta cgcctttgct tcctttgctt cttac 45 <210> 313 <211> 40 <212> DNA <213> CNAG_02476 3 flanking region primer 1 <400> 313 catggtcata gctgtttcct ggtgtcttga atccaatgcc 40 <210> 314 <211> 20 <212> DNA <213> CNAG_02476 3 flanking region primer 2 <400> 314 ctgctccata caaggcttag 20 <210> 315 <211> 20 <212> DNA <213> CNAG_02476 diagnostic screening primer, pairing with B79 <400> 315 tcgtcagcaa ttgaaaacct 20 <210> 316 <211> 19 <212> DNA <213> CNAG_02476 Southern blot probe primer <400> 316 atcaagtcag accacgagg 19 <210> 317 <211> 20 <212> DNA <213> NAT#219 STM primer <400> 317 ccctaaaacc ctacagcaat 20 <210> 318 <211> 24 <212> DNA <213> STM common primer <400> 318 gcatgccctg cccctaagaa ttcg 24 <210> 319 <211> 20 <212> DNA <213> CNAG_01841 5 flanking region primer 1 <400> 319 ccattatcat ccgtcaccac 20 <210> 320 <211> 37 <212> DNA <213> CNAG_01841 5 flanking region primer 2 <400> 320 ctggccgtcg ttttacgaag agggaagaga gggtaac 37 <210> 321 <211> 36 <212> DNA <213> CNAG_01841 3 flanking region primer 1 <400> 321 gtcatagctg tttcctgtga ggctgaggaa acagag 36 <210> 322 <211> 18 <212> DNA <213> CNAG_01841 3 flanking region primer 2 <400> 322 tccttttccc aacctgtc 18 <210> 323 <211> 21 <212> DNA <213> CNAG_01841 diagnostic screening primer, pairing with B79 <400> 323 ttcagtggtt ggtctctaca g 21 <210> 324 <211> 21 <212> DNA <213> CNAG_01841 Southern blot probe primer <400> 324 tggtaaaggt ctatcagtgc c 21 <210> 325 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 325 atgtaggtag ggtgataggt 20 <210> 326 <211> 24 <212> DNA <213> STM common primer <400> 326 gcatgccctg cccctaagaa ttcg 24 <210> 327 <211> 19 <212> DNA <213> CNAG_04457 5 flanking region primer 1 <400> 327 cttcaaaatc cagagcagg 19 <210> 328 <211> 38 <212> DNA <213> CNAG_04457 5 flanking region primer 2 <400> 328 ctggccgtcg ttttacctgg tctgatagtt cgtattcg 38 <210> 329 <211> 39 <212> DNA <213> CNAG_04457 3 flanking region primer 1 <400> 329 gtcatagctg tttcctggga cttgaacaat ggtatggac 39 <210> 330 <211> 19 <212> DNA <213> CNAG_04457 3 flanking region primer 2 <400> 330 tttagagcct acggtggtc 19 <210> 331 <211> 22 <212> DNA <213> CNAG_04457 diagnostic screening primer, pairing with B79 <400> 331 ttgcccgacg agatgctatg ac 22 <210> 332 <211> 20 <212> DNA <213> CNAG_04457 Southern blot probe primer <400> 332 tccgtctccg atgatttgcg 20 <210> 333 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 333 atgtaggtag ggtgataggt 20 <210> 334 <211> 24 <212> DNA <213> STM common primer <400> 334 gcatgccctg cccctaagaa ttcg 24 <210> 335 <211> 19 <212> DNA <213> CNAG_06252 5 flanking region primer 1 <400> 335 tctgacaggt tcaatcgcc 19 <210> 336 <211> 41 <212> DNA <213> CNAG_06252 5 flanking region primer 2 <400> 336 tcactggccg tcgttttacg catttctaca ccgtgattag c 41 <210> 337 <211> 40 <212> DNA <213> CNAG_06252 3 flanking region primer 1 <400> 337 catggtcata gctgtttcct gggatgtgcg gttttgttag 40 <210> 338 <211> 19 <212> DNA <213> CNAG_06252 3 flanking region primer 2 <400> 338 taacgaagtt ctccccgtg 19 <210> 339 <211> 20 <212> DNA <213> CNAG_06252 diagnostic screening primer, pairing with B79 <400> 339 tgagagggaa cgaaggatac 20 <210> 340 <211> 20 <212> DNA <213> CNAG_06252 Southern blot probe primer <400> 340 cttcgcctgt tggaaataag 20 <210> 341 <211> 20 <212> DNA <213> CNAG_06327 5 flanking region primer 1 <400> 341 ggagaaggga aagatgggtg 20 <210> 342 <211> 41 <212> DNA <213> CNAG_06327 5 flanking region primer 2 <400> 342 gctcactggc cgtcgtttta ctgtttccac tattgggaga g 41 <210> 343 <211> 40 <212> DNA <213> CNAG_06327 3 flanking region primer 1 <400> 343 catggtcata gctgtttcct gaggtgaagg tgtgagattg 40 <210> 344 <211> 18 <212> DNA <213> CNAG_06327 3 flanking region primer 2 <400> 344 caataatgtc ctcgccag 18 <210> 345 <211> 19 <212> DNA <213> CNAG_06327 diagnostic screening primer, pairing with B79 <400> 345 gtatgaaggg aaggtgctg 19 <210> 346 <211> 18 <212> DNA <213> CNAG_06327 Southern blot probe primer <400> 346 acgggttacg agaaatgc 18 <210> 347 <211> 19 <212> DNA <213> CNAG_03790 5 flanking region primer 1 <400> 347 tgagggttgt tgagggaag 19 <210> 348 <211> 40 <212> DNA <213> CNAG_03790 5 flanking region primer 2 <400> 348 gctcactggc cgtcgtttta cagcactgta tcgccgaaac 40 <210> 349 <211> 42 <212> DNA <213> CNAG_03790 3 flanking region primer 1 <400> 349 catggtcata gctgtttcct gctcgtagca atgtagatgt gc 42 <210> 350 <211> 21 <212> DNA <213> CNAG_03790 3 flanking region primer 2 <400> 350 tcagatgaag acgactacga c 21 <210> 351 <211> 19 <212> DNA <213> CNAG_03790 diagnostic screening primer, pairing with B79 <400> 351 cttccacaaa gccagattc 19 <210> 352 <211> 18 <212> DNA <213> CNAG_03790 Southern blot probe primer <400> 352 agattgcgtg aaacccac 18 <210> 353 <211> 19 <212> DNA <213> CNAG_05112 5 flanking region primer 1 <400> 353 atgtgcgaag tccaagagc 19 <210> 354 <211> 40 <212> DNA <213> CNAG_05112 5 flanking region primer 2 <400> 354 gctcactggc cgtcgtttta cgggtgaaga caacagatgg 40 <210> 355 <211> 41 <212> DNA <213> CNAG_05112 3 flanking region primer 1 <400> 355 catggtcata gctgtttcct gcagaccaac tggcgtaata g 41 <210> 356 <211> 19 <212> DNA <213> CNAG_05112 3 flanking region primer 2 <400> 356 acgaccaaat gctcttcgc 19 <210> 357 <211> 19 <212> DNA <213> CNAG_05112 diagnostic screening primer, pairing with B79 <400> 357 tcggatacag gcgatagag 19 <210> 358 <211> 19 <212> DNA <213> CNAG_05112 Southern blot probe primer <400> 358 tcaacagcag tttcaggtg 19 <210> 359 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 359 acagctccaa acctcgctaa acag 24 <210> 360 <211> 24 <212> DNA <213> STM common primer <400> 360 gcatgccctg cccctaagaa ttcg 24 <210> 361 <211> 19 <212> DNA <213> CNAG_00068 5 flanking region primer 1 <400> 361 tctatcacct ctgcgttgc 19 <210> 362 <211> 39 <212> DNA <213> CNAG_00068 5 flanking region primer 2 <400> 362 tcactggccg tcgttttaca ttcctaccac tatcccctg 39 <210> 363 <211> 40 <212> DNA <213> CNAG_00068 3 flanking region primer 1 <400> 363 catggtcata gctgtttcct gcggataaga tgcctgagac 40 <210> 364 <211> 22 <212> DNA <213> CNAG_00068 3 flanking region primer 2 <400> 364 ggtatgtttc tcggatgtaa gg 22 <210> 365 <211> 20 <212> DNA <213> CNAG_00068 diagnostic screening primer, pairing with B79 <400> 365 catcatcagg tatcagcagc 20 <210> 366 <211> 20 <212> DNA <213> CNAG_00068 Southern blot probe primer <400> 366 cttccaactt caatgttccc 20 <210> 367 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 367 cgcaaaatca ctagccctat agcg 24 <210> 368 <211> 24 <212> DNA <213> STM common primer <400> 368 gcatgccctg cccctaagaa ttcg 24 <210> 369 <211> 19 <212> DNA <213> CNAG_04184 5 flanking region primer 1 <400> 369 tcgttcacat aactgccac 19 <210> 370 <211> 38 <212> DNA <213> CNAG_04184 5 flanking region primer 2 <400> 370 tcactggccg tcgttttact cattgtgagc ctcgtatg 38 <210> 371 <211> 39 <212> DNA <213> CNAG_04184 3 flanking region primer 1 <400> 371 catggtcata gctgtttcct gaatggtgac ggttcaagg 39 <210> 372 <211> 18 <212> DNA <213> CNAG_04184 3 flanking region primer 2 <400> 372 cttttccact gaatccgc 18 <210> 373 <211> 19 <212> DNA <213> CNAG_04184 diagnostic screening primer, pairing with B79 <400> 373 tggtccgatg ccgtaaatc 19 <210> 374 <211> 19 <212> DNA <213> CNAG_04184 Southern blot probe primer <400> 374 caagggaatg actcactcc 19 <210> 375 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 375 ccatagcgat atctacccca atct 24 <210> 376 <211> 24 <212> DNA <213> STM common primer <400> 376 gcatgccctg cccctaagaa ttcg 24 <210> 377 <211> 18 <212> DNA <213> CNAG_03018 5 flanking region primer 1 <400> 377 agccgtttgt cattctgc 18 <210> 378 <211> 37 <212> DNA <213> CNAG_03018 5 flanking region primer 2 <400> 378 tcactggccg tcgttttacg tcgttgctca ttttgcg 37 <210> 379 <211> 39 <212> DNA <213> CNAG_03018 3 flanking region primer 1 <400> 379 catggtcata gctgtttcct ggcgaacctc caataaagc 39 <210> 380 <211> 18 <212> DNA <213> CNAG_03018 3 flanking region primer 2 <400> 380 cccttttttc cctgcttc 18 <210> 381 <211> 18 <212> DNA <213> CNAG_03018 diagnostic screening primer, pairing with B79 <400> 381 aatagggatg ccaaccag 18 <210> 382 <211> 18 <212> DNA <213> CNAG_03018 Southern blot probe primer <400> 382 caaaaagtgt cagcacgc 18 <210> 383 <211> 20 <212> DNA <213> NAT#150 STM primer <400> 383 acatacaccc ccatcccccc 20 <210> 384 <211> 24 <212> DNA <213> STM common primer <400> 384 gcatgccctg cccctaagaa ttcg 24 <210> 385 <211> 20 <212> DNA <213> CNAG_01708 5 flanking region primer 1 <400> 385 ccgactcgtt cgtctatctg 20 <210> 386 <211> 41 <212> DNA <213> CNAG_01708 5 flanking region primer 2 <400> 386 tcactggccg tcgttttact ggagaggata gatgtgatga c 41 <210> 387 <211> 41 <212> DNA <213> CNAG_01708 3 flanking region primer 1 <400> 387 catggtcata gctgtttcct gaacttctac ccaggtcgtg c 41 <210> 388 <211> 18 <212> DNA <213> CNAG_01708 3 flanking region primer 2 <400> 388 aactgccctt cttcaagc 18 <210> 389 <211> 20 <212> DNA <213> CNAG_01708 diagnostic screening primer, pairing with B79 <400> 389 tagcctctcc caatgtccac 20 <210> 390 <211> 20 <212> DNA <213> CNAG_01708 Southern blot probe primer <400> 390 cgattctcct ttgctgagtc 20 <210> 391 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 391 acgcaccaga cacacaacca g 21 <210> 392 <211> 24 <212> DNA <213> STM common primer <400> 392 gcatgccctg cccctaagaa ttcg 24 <210> 393 <211> 19 <212> DNA <213> CNAG_05861 5 flanking region primer 1 <400> 393 ggttttggtc acttcaccg 19 <210> 394 <211> 38 <212> DNA <213> CNAG_05861 5 flanking region primer 2 <400> 394 tcactggccg tcgttttaca gcagaaagga gatgatgc 38 <210> 395 <211> 40 <212> DNA <213> CNAG_05861 3 flanking region primer 1 <400> 395 catggtcata gctgtttcct ggacgacttt tacgaccagg 40 <210> 396 <211> 18 <212> DNA <213> CNAG_05861 3 flanking region primer 2 <400> 396 aaatggaaca agcccgtc 18 <210> 397 <211> 18 <212> DNA <213> CNAG_05861 diagnostic screening primer, pairing with B79 <400> 397 tctgaagggc gtaaatgc 18 <210> 398 <211> 18 <212> DNA <213> CNAG_05861 Southern blot probe primer <400> 398 aaggaatggg ttgttggg 18 <210> 399 <211> 24 <212> DNA <213> NAT#184 STM primer <400> 399 atatatggct cgagctagat agag 24 <210> 400 <211> 24 <212> DNA <213> STM common primer <400> 400 gcatgccctg cccctaagaa ttcg 24 <210> 401 <211> 20 <212> DNA <213> CNAG_02066 5 flanking region primer 1 <400> 401 ccccattgta aacattggtg 20 <210> 402 <211> 36 <212> DNA <213> CNAG_02066 5 flanking region primer 2 <400> 402 ctggccgtcg ttttaccgtg atgtggagtt gattcg 36 <210> 403 <211> 36 <212> DNA <213> CNAG_02066 3 flanking region primer 1 <400> 403 gtcatagctg tttcctgtgc tgccaatgga aggttc 36 <210> 404 <211> 20 <212> DNA <213> CNAG_02066 3 flanking region primer 2 <400> 404 atagtctcgc tccttggtgc 20 <210> 405 <211> 19 <212> DNA <213> CNAG_02066 diagnostic screening primer, pairing with B79 <400> 405 gttcgttatc cgctctttg 19 <210> 406 <211> 19 <212> DNA <213> CNAG_02066 Southern blot probe primer <400> 406 acataggcga gacagttgc 19 <210> 407 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 407 atatggatgt ttttagcgag 20 <210> 408 <211> 24 <212> DNA <213> STM common primer <400> 408 gcatgccctg cccctaagaa ttcg 24 <210> 409 <211> 19 <212> DNA <213> CNAG_03527 5 flanking region primer 1 <400> 409 ccatctcacg gttgatacg 19 <210> 410 <211> 35 <212> DNA <213> CNAG_03527 5 flanking region primer 2 <400> 410 ctggccgtcg ttttactcaa acccttatcc caccg 35 <210> 411 <211> 37 <212> DNA <213> CNAG_03527 3 flanking region primer 1 <400> 411 gtcatagctg tttcctgtct ttccctgctt tgcctgc 37 <210> 412 <211> 20 <212> DNA <213> CNAG_03527 3 flanking region primer 2 <400> 412 cgtcatccgt cgtgtaactg 20 <210> 413 <211> 19 <212> DNA <213> CNAG_03527 diagnostic screening primer, pairing with B79 <400> 413 tctcgcaagg tcagagttc 19 <210> 414 <211> 21 <212> DNA <213> CNAG_03527 Southern blot probe primer <400> 414 ctatgagcga cattcacaga c 21 <210> 415 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 415 gatctctcgc gcttggggga 20 <210> 416 <211> 24 <212> DNA <213> STM common primer <400> 416 gcatgccctg cccctaagaa ttcg 24 <210> 417 <211> 18 <212> DNA <213> CNAG_04263 5 flanking region primer 1 <400> 417 tgtttccaca caccgatg 18 <210> 418 <211> 36 <212> DNA <213> CNAG_04263 5 flanking region primer 2 <400> 418 ctggccgtcg ttttacggtg gtggtcatct gtatgc 36 <210> 419 <211> 37 <212> DNA <213> CNAG_04263 3 flanking region primer 1 <400> 419 gtcatagctg tttcctgaga agatggagac gcacacg 37 <210> 420 <211> 19 <212> DNA <213> CNAG_04263 3 flanking region primer 2 <400> 420 gctattgcgt gaagactcc 19 <210> 421 <211> 20 <212> DNA <213> CNAG_04263 diagnostic screening primer, pairing with B79 <400> 421 cgtgattgag agtagcaacc 20 <210> 422 <211> 19 <212> DNA <213> CNAG_04263 Southern blot probe primer <400> 422 gtaatgtcag tgagtgcgg 19 <210> 423 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 423 tatccccctc tccgctctct agca 24 <210> 424 <211> 24 <212> DNA <213> STM common primer <400> 424 gcatgccctg cccctaagaa ttcg 24 <210> 425 <211> 20 <212> DNA <213> CNAG_00018 5 flanking region primer 1 <400> 425 tttgcctcaa tctctctcag 20 <210> 426 <211> 41 <212> DNA <213> CNAG_00018 5 flanking region primer 2 <400> 426 gctcactggc cgtcgtttta cggaatacaa catccacctc c 41 <210> 427 <211> 42 <212> DNA <213> CNAG_00018 3 flanking region primer 1 <400> 427 catggtcata gctgtttcct ggcacgggtt acattctaca tc 42 <210> 428 <211> 20 <212> DNA <213> CNAG_00018 3 flanking region primer 2 <400> 428 ggggaggact tgaacttgag 20 <210> 429 <211> 18 <212> DNA <213> CNAG_00018 diagnostic screening primer, pairing with B79 <400> 429 aacaaactgc cttcctcc 18 <210> 430 <211> 19 <212> DNA <213> CNAG_00018 Southern blot probe primer <400> 430 gcttggtcgg ataaagatg 19 <210> 431 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 431 gcggtcgctt tatagcgatt 20 <210> 432 <211> 24 <212> DNA <213> STM common primer <400> 432 gcatgccctg cccctaagaa ttcg 24 <210> 433 <211> 19 <212> DNA <213> CNAG_05176 5 flanking region primer 1 <400> 433 tgagaaggag gatggtcac 19 <210> 434 <211> 37 <212> DNA <213> CNAG_05176 5 flanking region primer 2 <400> 434 tcactggccg tcgttttaca tgtggatagt gttgcgg 37 <210> 435 <211> 40 <212> DNA <213> CNAG_05176 3 flanking region primer 1 <400> 435 catggtcata gctgtttcct ggcaaaaagg gagaatgtcg 40 <210> 436 <211> 19 <212> DNA <213> CNAG_05176 3 flanking region primer 2 <400> 436 tcatacagcc gagttgacc 19 <210> 437 <211> 19 <212> DNA <213> CNAG_05176 diagnostic screening primer, pairing with B79 <400> 437 tgtgagtatg cctatcccg 19 <210> 438 <211> 19 <212> DNA <213> CNAG_05176 Southern blot probe primer <400> 438 tcaccgagac taatggcac 19 <210> 439 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 439 gcggtcgctt tatagcgatt 20 <210> 440 <211> 24 <212> DNA <213> STM common primer <400> 440 gcatgccctg cccctaagaa ttcg 24 <210> 441 <211> 19 <212> DNA <213> CNAG_06339 5 flanking region primer 1 <400> 441 tagtgaaaga ctcggcacc 19 <210> 442 <211> 37 <212> DNA <213> CNAG_06339 5 flanking region primer 2 <400> 442 tcactggccg tcgttttacc agtcaccgca aagattc 37 <210> 443 <211> 41 <212> DNA <213> CNAG_06339 3 flanking region primer 1 <400> 443 catggtcata gctgtttcct gcgcttcgtt attacctgtt c 41 <210> 444 <211> 22 <212> DNA <213> CNAG_06339 3 flanking region primer 2 <400> 444 ctccaggata agtagtgtga gg 22 <210> 445 <211> 20 <212> DNA <213> CNAG_06339 diagnostic screening primer, pairing with B79 <400> 445 gggaaagtag ggtaaccacc 20 <210> 446 <211> 20 <212> DNA <213> CNAG_06339 Southern blot probe primer <400> 446 gaggagaggt gaataatgcc 20 <210> 447 <211> 24 <212> DNA <213> NAT#213 STM primer <400> 447 ctggggattt tgatgtgtct atgt 24 <210> 448 <211> 24 <212> DNA <213> STM common primer <400> 448 gcatgccctg cccctaagaa ttcg 24 <210> 449 <211> 19 <212> DNA <213> CNAG_07593 5 flanking region primer 1 <400> 449 gattgcttgg atggacttc 19 <210> 450 <211> 39 <212> DNA <213> CNAG_07593 5 flanking region primer 2 <400> 450 tcactggccg tcgttttact gcctttcttg ccgaactcg 39 <210> 451 <211> 40 <212> DNA <213> CNAG_07593 3 flanking region primer 1 <400> 451 catggtcata gctgtttcct gtttgaagga ccaggagcag 40 <210> 452 <211> 19 <212> DNA <213> CNAG_07593 3 flanking region primer 2 <400> 452 tcaagtgaca cagtgccac 19 <210> 453 <211> 19 <212> DNA <213> CNAG_07593 diagnostic screening primer, pairing with B79 <400> 453 ccataatcag acagcgttg 19 <210> 454 <211> 19 <212> DNA <213> CNAG_07593 Southern blot probe primer <400> 454 ggtttggggg aatagtgag 19 <210> 455 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 455 ctccacatcc atcgctccaa 20 <210> 456 <211> 24 <212> DNA <213> STM common primer <400> 456 gcatgccctg cccctaagaa ttcg 24 <210> 457 <211> 20 <212> DNA <213> CNAG_00670 5 flanking region primer 1 <400> 457 tgatgaagat gctgatgagg 20 <210> 458 <211> 41 <212> DNA <213> CNAG_00670 5 flanking region primer 2 <400> 458 gctcactggc cgtcgtttta ctcatttggc gtctcactta c 41 <210> 459 <211> 39 <212> DNA <213> CNAG_00670 3 flanking region primer 1 <400> 459 catggtcata gctgtttcct gtgtgaaagg ggagtgatg 39 <210> 460 <211> 19 <212> DNA <213> CNAG_00670 3 flanking region primer 2 <400> 460 agaatacatc gtggttggc 19 <210> 461 <211> 22 <212> DNA <213> CNAG_00670 diagnostic screening primer, pairing with B79 <400> 461 ggaagtgtag agtcaggaga tg 22 <210> 462 <211> 21 <212> DNA <213> CNAG_00670 Southern blot probe primer <400> 462 ggatgaaagg tgaaaggaca c 21 <210> 463 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 463 aacctttaaa tgggtagag 19 <210> 464 <211> 24 <212> DNA <213> STM common primer <400> 464 gcatgccctg cccctaagaa ttcg 24 <210> 465 <211> 18 <212> DNA <213> CNAG_05010 5 flanking region primer 1 <400> 465 tccttccttt ctaagccg 18 <210> 466 <211> 41 <212> DNA <213> CNAG_05010 5 flanking region primer 2 <400> 466 gctcactggc cgtcgtttta ctgtccccta cttttactcc g 41 <210> 467 <211> 41 <212> DNA <213> CNAG_05010 3 flanking region primer 1 <400> 467 catggtcata gctgtttcct gtgagggttt gataccttgt g 41 <210> 468 <211> 18 <212> DNA <213> CNAG_05010 3 flanking region primer 2 <400> 468 aagagcaaca tccgcaag 18 <210> 469 <211> 19 <212> DNA <213> CNAG_05010 diagnostic screening primer, pairing with B79 <400> 469 aacagccttg ccttacagc 19 <210> 470 <211> 19 <212> DNA <213> CNAG_05010 Southern blot probe primer <400> 470 agtggtgaag ggctaaatg 19 <210> 471 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 471 aacctttaaa tgggtagag 19 <210> 472 <211> 24 <212> DNA <213> STM common primer <400> 472 gcatgccctg cccctaagaa ttcg 24 <210> 473 <211> 18 <212> DNA <213> CNAG_04345 5 flanking region primer 1 <400> 473 aaaaggcagt cgtcttcg 18 <210> 474 <211> 40 <212> DNA <213> CNAG_04345 5 flanking region primer 2 <400> 474 ctggccgtcg ttttaccctt ttagatactc acacaagccc 40 <210> 475 <211> 36 <212> DNA <213> CNAG_04345 3 flanking region primer 1 <400> 475 gtcatagctg tttcctgttc tgggatgact tcacgc 36 <210> 476 <211> 19 <212> DNA <213> CNAG_04345 3 flanking region primer 2 <400> 476 ttactgctcc tggtttggg 19 <210> 477 <211> 18 <212> DNA <213> CNAG_04345 diagnostic screening primer, pairing with B79 <400> 477 cctgcttcac aaaatggc 18 <210> 478 <211> 22 <212> DNA <213> CNAG_04345 Southern blot probe primer <400> 478 gagtagtggt gtcagtgtga tg 22 <210> 479 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 479 ccatagaact agctaaagca 20 <210> 480 <211> 24 <212> DNA <213> STM common primer <400> 480 gcatgccctg cccctaagaa ttcg 24 <210> 481 <211> 19 <212> DNA <213> CNAG_02774 5 flanking region primer 1 <400> 481 ttgagccagc ctaagcaag 19 <210> 482 <211> 35 <212> DNA <213> CNAG_02774 5 flanking region primer 2 <400> 482 ctggccgtcg ttttactctt cttttctgcg gtcac 35 <210> 483 <211> 36 <212> DNA <213> CNAG_02774 3 flanking region primer 1 <400> 483 gtcatagctg tttcctggct tgatttcttg gaccac 36 <210> 484 <211> 19 <212> DNA <213> CNAG_02774 3 flanking region primer 2 <400> 484 cgagataaga agcaacgcc 19 <210> 485 <211> 19 <212> DNA <213> CNAG_02774 diagnostic screening primer, pairing with B79 <400> 485 tgccagccaa cctaatctg 19 <210> 486 <211> 20 <212> DNA <213> CNAG_02774 Southern blot probe primer <400> 486 gagaggcagg gagataactc 20 <210> 487 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 487 atgtaggtag ggtgataggt 20 <210> 488 <211> 24 <212> DNA <213> STM common primer <400> 488 gcatgccctg cccctaagaa ttcg 24 <210> 489 <211> 20 <212> DNA <213> CNAG_06425 5 flanking region primer 1 <400> 489 ctttcacgaa cgcttgattc 20 <210> 490 <211> 35 <212> DNA <213> CNAG_06425 5 flanking region primer 2 <400> 490 ctggccgtcg ttttacttgg acagtttttc ggcgg 35 <210> 491 <211> 36 <212> DNA <213> CNAG_06425 3 flanking region primer 1 <400> 491 gtcatagctg tttcctgatg tgcctgcgga tttctc 36 <210> 492 <211> 19 <212> DNA <213> CNAG_06425 3 flanking region primer 2 <400> 492 tgagaccgct gaaagttcc 19 <210> 493 <211> 19 <212> DNA <213> CNAG_06425 diagnostic screening primer, pairing with B79 <400> 493 taatcgccga caaagggtg 19 <210> 494 <211> 18 <212> DNA <213> CNAG_06425 Southern blot probe primer <400> 494 cccttttttt gtgcgacc 18 <210> 495 <211> 24 <212> DNA <213> NAT#288 STM primer <400> 495 ctatccaact agacctctag ctac 24 <210> 496 <211> 24 <212> DNA <213> STM common primer <400> 496 gcatgccctg cccctaagaa ttcg 24 <210> 497 <211> 22 <212> DNA <213> CNAG_05255 5 flanking region primer 1 <400> 497 atggtcaccg tcacaaggaa gc 22 <210> 498 <211> 40 <212> DNA <213> CNAG_05255 5 flanking region primer 2 <400> 498 ctggccgtcg ttttactaca cattcggctt ggttgcggac 40 <210> 499 <211> 38 <212> DNA <213> CNAG_05255 3 flanking region primer 1 <400> 499 gtcatagctg tttcctgcga ggtttgtatc ccagtcag 38 <210> 500 <211> 20 <212> DNA <213> CNAG_05255 3 flanking region primer 2 <400> 500 aggggctatc tctgcttcac 20 <210> 501 <211> 24 <212> DNA <213> CNAG_05255 diagnostic screening primer, pairing with B79 <400> 501 ttctgtgccc acctctactc tcag 24 <210> 502 <211> 20 <212> DNA <213> CNAG_05255 Southern blot probe primer <400> 502 accaaaactc gtcgtaggcg 20 <210> 503 <211> 24 <212> DNA <213> NAT#288 STM primer <400> 503 ctatccaact agacctctag ctac 24 <210> 504 <211> 24 <212> DNA <213> STM common primer <400> 504 gcatgccctg cccctaagaa ttcg 24 <210> 505 <211> 19 <212> DNA <213> CNAG_05049 5 flanking region primer 1 <400> 505 atcaggacga gtggcattg 19 <210> 506 <211> 39 <212> DNA <213> CNAG_05049 5 flanking region primer 2 <400> 506 tcactggccg tcgttttacg agtcttcttt gggcatagc 39 <210> 507 <211> 40 <212> DNA <213> CNAG_05049 3 flanking region primer 1 <400> 507 catggtcata gctgtttcct gagtggatga tgaggactcg 40 <210> 508 <211> 21 <212> DNA <213> CNAG_05049 3 flanking region primer 2 <400> 508 gtttcctatc ttcagtcgga g 21 <210> 509 <211> 20 <212> DNA <213> CNAG_05049 diagnostic screening primer, pairing with B79 <400> 509 gatagcagag gagaaaaggc 20 <210> 510 <211> 18 <212> DNA <213> CNAG_05049 Southern blot probe primer <400> 510 tatttggggt tgaggctg 18 <210> 511 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 511 ctatcgacca accaacacag 20 <210> 512 <211> 24 <212> DNA <213> STM common primer <400> 512 gcatgccctg cccctaagaa ttcg 24 <210> 513 <211> 18 <212> DNA <213> CNAG_05222 5 flanking region primer 1 <400> 513 gagggtcctt tggttttg 18 <210> 514 <211> 37 <212> DNA <213> CNAG_05222 5 flanking region primer 2 <400> 514 tcactggccg tcgttttact ttcacactca cggttgg 37 <210> 515 <211> 39 <212> DNA <213> CNAG_05222 3 flanking region primer 1 <400> 515 catggtcata gctgtttcct ggttgcccct tttgacaag 39 <210> 516 <211> 19 <212> DNA <213> CNAG_05222 3 flanking region primer 2 <400> 516 aacttacggc gacaacaac 19 <210> 517 <211> 18 <212> DNA <213> CNAG_05222 diagnostic screening primer, pairing with B79 <400> 517 tccacataat agccaccg 18 <210> 518 <211> 20 <212> DNA <213> CNAG_05222 Southern blot probe primer <400> 518 gctatgttca ggagcgatac 20 <210> 519 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 519 ctatcgacca accaacacag 20 <210> 520 <211> 24 <212> DNA <213> STM common primer <400> 520 gcatgccctg cccctaagaa ttcg 24 <210> 521 <211> 18 <212> DNA <213> CNAG_04130 5 flanking region primer 1 <400> 521 cattgacgcc atagcaag 18 <210> 522 <211> 38 <212> DNA <213> CNAG_04130 5 flanking region primer 2 <400> 522 tcactggccg tcgttttacg tgtttggaat gcggtttc 38 <210> 523 <211> 41 <212> DNA <213> CNAG_04130 3 flanking region primer 1 <400> 523 catggtcata gctgtttcct gtggatgagg ttgaggggaa c 41 <210> 524 <211> 21 <212> DNA <213> CNAG_04130 3 flanking region primer 2 <400> 524 gtagtgtaag ggcaggaaga g 21 <210> 525 <211> 21 <212> DNA <213> CNAG_04130 diagnostic screening primer, pairing with B79 <400> 525 ttctactcat aagggctacg c 21 <210> 526 <211> 21 <212> DNA <213> CNAG_04130 Southern blot probe primer <400> 526 ttctactcat aagggctacg c 21 <210> 527 <211> 19 <212> DNA <213> CNAG_00193 5 flanking region primer 1 <400> 527 gggcagaaat agttggcag 19 <210> 528 <211> 34 <212> DNA <213> CNAG_00193 5 flanking region primer 2 <400> 528 ctggccgtcg ttttactttt tccccccgat gtag 34 <210> 529 <211> 36 <212> DNA <213> CNAG_00193 3 flanking region primer 1 <400> 529 gtcatagctg tttcctgttc ccgcaagaac agtgtc 36 <210> 530 <211> 20 <212> DNA <213> CNAG_00193 3 flanking region primer 2 <400> 530 ttcaggatgt ccaaactcag 20 <210> 531 <211> 19 <212> DNA <213> CNAG_00193 diagnostic screening primer, pairing with B79 <400> 531 aaggggtaag aaatcgggc 19 <210> 532 <211> 19 <212> DNA <213> CNAG_00193 Southern blot probe primer <400> 532 tgctgagaga cagggaatc 19 <210> 533 <211> 20 <212> DNA <213> NAT#227 STM primer <400> 533 tcgtggttta gagggagcgc 20 <210> 534 <211> 24 <212> DNA <213> STM common primer <400> 534 gcatgccctg cccctaagaa ttcg 24 <210> 535 <211> 20 <212> DNA <213> CNAG_02700 5 flanking region primer 1 <400> 535 tgaaatcaga ctccagcctc 20 <210> 536 <211> 38 <212> DNA <213> CNAG_02700 5 flanking region primer 2 <400> 536 tcactggccg tcgttttacg cgaccacatt tacactcg 38 <210> 537 <211> 41 <212> DNA <213> CNAG_02700 3 flanking region primer 1 <400> 537 catggtcata gctgtttcct ggccacagaa caataactgg g 41 <210> 538 <211> 19 <212> DNA <213> CNAG_02700 3 flanking region primer 2 <400> 538 gcgtatgaat aagcgttgg 19 <210> 539 <211> 18 <212> DNA <213> CNAG_02700 diagnostic screening primer, pairing with B79 <400> 539 agcaaaacag aagcagcg 18 <210> 540 <211> 18 <212> DNA <213> CNAG_02700 Southern blot probe primer <400> 540 agaaagcagc atttgggg 18 <210> 541 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 541 atgtaggtag ggtgataggt 20 <210> 542 <211> 24 <212> DNA <213> STM common primer <400> 542 gcatgccctg cccctaagaa ttcg 24 <210> 543 <211> 19 <212> DNA <213> CNAG_04586 5 flanking region primer 1 <400> 543 tccatctccc aatccacag 19 <210> 544 <211> 38 <212> DNA <213> CNAG_04586 5 flanking region primer 2 <400> 544 tcactggccg tcgttttacg cttgattagg cggataag 38 <210> 545 <211> 41 <212> DNA <213> CNAG_04586 3 flanking region primer 1 <400> 545 catggtcata gctgtttcct gtggacgaaa agacccctac c 41 <210> 546 <211> 18 <212> DNA <213> CNAG_04586 3 flanking region primer 2 <400> 546 aacactggaa tgtgcgtg 18 <210> 547 <211> 20 <212> DNA <213> CNAG_04586 diagnostic screening primer, pairing with B79 <400> 547 ttggtggtgc ggttttgtgc 20 <210> 548 <211> 18 <212> DNA <213> CNAG_04586 Southern blot probe primer <400> 548 tgatgttggt tcgtgacc 18 <210> 549 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 549 acgcaccaga cacacaacca g 21 <210> 550 <211> 24 <212> DNA <213> STM common primer <400> 550 gcatgccctg cccctaagaa ttcg 24 <210> 551 <211> 18 <212> DNA <213> CNAG_05153 5 flanking region primer 1 <400> 551 aaaaccaagt ggaacccg 18 <210> 552 <211> 35 <212> DNA <213> CNAG_05153 5 flanking region primer 2 <400> 552 ctggccgtcg ttttactggt tgacgctgtt agcac 35 <210> 553 <211> 37 <212> DNA <213> CNAG_05153 3 flanking region primer 1 <400> 553 gtcatagctg tttcctgata ggcaggctgg tggaaac 37 <210> 554 <211> 19 <212> DNA <213> CNAG_05153 3 flanking region primer 2 <400> 554 accacatttg cttccgagc 19 <210> 555 <211> 21 <212> DNA <213> CNAG_05153 diagnostic screening primer, pairing with JOHE12579 <400> 555 tggctgaact gtaggttagt g 21 <210> 556 <211> 18 <212> DNA <213> CNAG_05153 Southern blot probe primer <400> 556 cggcgataag gtttttgc 18 <210> 557 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 557 accgacagct cgaacaagca agag 24 <210> 558 <211> 24 <212> DNA <213> STM common primer <400> 558 gcatgccctg cccctaagaa ttcg 24 <210> 559 <211> 20 <212> DNA <213> CNAG_02723 5 flanking region primer 1 <400> 559 gcttccttat cggtagggac 20 <210> 560 <211> 39 <212> DNA <213> CNAG_02723 5 flanking region primer 2 <400> 560 tcactggccg tcgttttacg actggacatt gacgaagac 39 <210> 561 <211> 40 <212> DNA <213> CNAG_02723 3 flanking region primer 1 <400> 561 catggtcata gctgtttcct gtggtgacag tgacgcttac 40 <210> 562 <211> 19 <212> DNA <213> CNAG_02723 3 flanking region primer 2 <400> 562 attgtcagga tgtgggttc 19 <210> 563 <211> 21 <212> DNA <213> CNAG_02723 diagnostic screening primer, pairing with B79 <400> 563 gcttactccc aaactcacac c 21 <210> 564 <211> 18 <212> DNA <213> CNAG_02723 Southern blot probe primer <400> 564 atggatttgg cagaacgg 18 <210> 565 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 565 caccctctat ctcgagaaag ctcc 24 <210> 566 <211> 24 <212> DNA <213> STM common primer <400> 566 gcatgccctg cccctaagaa ttcg 24 <210> 567 <211> 19 <212> DNA <213> CNAG_03741 5 flanking region primer 1 <400> 567 gccgtctcct gatgtaatc 19 <210> 568 <211> 36 <212> DNA <213> CNAG_03741 5 flanking region primer 2 <400> 568 ctggccgtcg ttttaccgac tgtcatcaac gactgg 36 <210> 569 <211> 36 <212> DNA <213> CNAG_03741 3 flanking region primer 1 <400> 569 ctggccgtcg ttttacgcgt caacaggaat aaaacc 36 <210> 570 <211> 22 <212> DNA <213> CNAG_03741 3 flanking region primer 2 <400> 570 ctgtgccttt ctccaagata ag 22 <210> 571 <211> 22 <212> DNA <213> CNAG_03741 diagnostic screening primer, pairing with B79 <400> 571 ggataagcag aacatagttg cg 22 <210> 572 <211> 21 <212> DNA <213> CNAG_03741 Southern blot probe primer <400> 572 tctcttcttc ttctgacctg c 21 <210> 573 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 573 gagatctttc gggaggtctg gatt 24 <210> 574 <211> 24 <212> DNA <213> STM common primer <400> 574 gcatgccctg cccctaagaa ttcg 24 <210> 575 <211> 18 <212> DNA <213> CNAG_07411 5 flanking region primer 1 <400> 575 agacccgcca aaacaatc 18 <210> 576 <211> 35 <212> DNA <213> CNAG_07411 5 flanking region primer 2 <400> 576 ctggccgtcg ttttacttgg acctacgctg acttc 35 <210> 577 <211> 37 <212> DNA <213> CNAG_07411 3 flanking region primer 1 <400> 577 gtcatagctg tttcctggac ttgcgttctg attttgg 37 <210> 578 <211> 20 <212> DNA <213> CNAG_07411 3 flanking region primer 2 <400> 578 gctgttatca agagaatgcc 20 <210> 579 <211> 19 <212> DNA <213> CNAG_07411 diagnostic screening primer, pairing with B79 <400> 579 acgactggct atcaagcag 19 <210> 580 <211> 19 <212> DNA <213> CNAG_07411 Southern blot probe primer <400> 580 tacgaggatt gagtctggc 19 <210> 581 <211> 24 <212> DNA <213> NAT#288 STM primer <400> 581 ctatccaact agacctctag ctac 24 <210> 582 <211> 24 <212> DNA <213> STM common primer <400> 582 gcatgccctg cccctaagaa ttcg 24 <210> 583 <211> 20 <212> DNA <213> CNAG_03849 5 flanking region primer 1 <400> 583 tagtggttcc cgtttgtgcg 20 <210> 584 <211> 40 <212> DNA <213> CNAG_03849 5 flanking region primer 2 <400> 584 gctcactggc cgtcgtttta ccttacgcct ttttccatcg 40 <210> 585 <211> 41 <212> DNA <213> CNAG_03849 3 flanking region primer 1 <400> 585 catggtcata gctgtttcct gtcaaacggc gattggtggt c 41 <210> 586 <211> 20 <212> DNA <213> CNAG_03849 3 flanking region primer 2 <400> 586 tggtgaaggg gaagattggg 20 <210> 587 <211> 19 <212> DNA <213> CNAG_03849 diagnostic screening primer, pairing with B79 <400> 587 aacaagccga ggaatgagg 19 <210> 588 <211> 19 <212> DNA <213> CNAG_03849 Southern blot probe primer <400> 588 tgagtgaacg aagagtgcc 19 <210> 589 <211> 20 <212> DNA <213> NAT#6 STM primer <400> 589 atagctacca cacgatagct 20 <210> 590 <211> 24 <212> DNA <213> STM common primer <400> 590 gcatgccctg cccctaagaa ttcg 24 <210> 591 <211> 19 <212> DNA <213> CNAG_04916 5 flanking region primer 1 <400> 591 ttgcgttgag acatcactg 19 <210> 592 <211> 43 <212> DNA <213> CNAG_04916 5 flanking region primer 2 <400> 592 gctcactggc cgtcgtttta cgctcctatg ttaccaatgt tgc 43 <210> 593 <211> 40 <212> DNA <213> CNAG_04916 3 flanking region primer 1 <400> 593 catggtcata gctgtttcct gacccatttg ctttcctggc 40 <210> 594 <211> 20 <212> DNA <213> CNAG_04916 3 flanking region primer 2 <400> 594 tctcaacgag tcgctttcgc 20 <210> 595 <211> 20 <212> DNA <213> CNAG_04916 diagnostic screening primer, pairing with B79 <400> 595 aaaagcatcc tgcccagcac 20 <210> 596 <211> 20 <212> DNA <213> CNAG_04916 Southern blot probe primer <400> 596 gttgaaagta gttgcgttgg 20 <210> 597 <211> 20 <212> DNA <213> NAT#212 STM primer <400> 597 agagcgatcg cgttatagat 20 <210> 598 <211> 24 <212> DNA <213> STM common primer <400> 598 gcatgccctg cccctaagaa ttcg 24 <210> 599 <211> 19 <212> DNA <213> CNAG_04012 5 flanking region primer 1 <400> 599 ttggtggagt gattgaccc 19 <210> 600 <211> 36 <212> DNA <213> CNAG_04012 5 flanking region primer 2 <400> 600 ctggccgtcg ttttacaatc aaaccagaag gagagc 36 <210> 601 <211> 39 <212> DNA <213> CNAG_04012 3 flanking region primer 1 <400> 601 gtcatagctg tttcctgagg aggataaagg aagagagtg 39 <210> 602 <211> 19 <212> DNA <213> CNAG_04012 3 flanking region primer 2 <400> 602 ttatgctggt ggtgcctac 19 <210> 603 <211> 19 <212> DNA <213> CNAG_04012 diagnostic screening primer, pairing with B79 <400> 603 accgaagagg atgaggttg 19 <210> 604 <211> 20 <212> DNA <213> CNAG_04012 Southern blot probe primer <400> 604 cagtggtggt tgagattgac 20 <210> 605 <211> 20 <212> DNA <213> NAT#212 STM primer <400> 605 agagcgatcg cgttatagat 20 <210> 606 <211> 24 <212> DNA <213> STM common primer <400> 606 gcatgccctg cccctaagaa ttcg 24 <210> 607 <211> 20 <212> DNA <213> CNAG_04090 5 flanking region primer 1 <400> 607 gagtagagga gtggattggg 20 <210> 608 <211> 39 <212> DNA <213> CNAG_04090 5 flanking region primer 2 <400> 608 gctcactggc cgtcgtttta cccattgttc aagcagagc 39 <210> 609 <211> 42 <212> DNA <213> CNAG_04090 3 flanking region primer 1 <400> 609 catggtcata gctgtttcct gggcttactg ctatcttgat gc 42 <210> 610 <211> 24 <212> DNA <213> CNAG_04090 3 flanking region primer 2 <400> 610 gaaagaggaa ctacatactg ggtc 24 <210> 611 <211> 20 <212> DNA <213> CNAG_04090 diagnostic screening primer, pairing with B79 <400> 611 gtcttttgct ccttgaaacc 20 <210> 612 <211> 20 <212> DNA <213> CNAG_04090 Southern blot probe primer <400> 612 accaggtgta aacccagtcc 20 <210> 613 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 613 cagatctcga acgataccca 20 <210> 614 <211> 24 <212> DNA <213> STM common primer <400> 614 gcatgccctg cccctaagaa ttcg 24 <210> 615 <211> 21 <212> DNA <213> CNAG_05375 5 flanking region primer 1 <400> 615 ggaagcatta ctgtcccata g 21 <210> 616 <211> 43 <212> DNA <213> CNAG_05375 5 flanking region primer 2 <400> 616 gctcactggc cgtcgtttta cagaaacagg agagagagta gcc 43 <210> 617 <211> 39 <212> DNA <213> CNAG_05375 3 flanking region primer 1 <400> 617 catggtcata gctgtttcct gtcttggatt tcaacgggg 39 <210> 618 <211> 20 <212> DNA <213> CNAG_05375 3 flanking region primer 2 <400> 618 gtagttccgt tatttgctgg 20 <210> 619 <211> 22 <212> DNA <213> CNAG_05375 diagnostic screening primer, pairing with B79 <400> 619 ggttgaagat ggacgagtag ac 22 <210> 620 <211> 20 <212> DNA <213> CNAG_05375 Southern blot probe primer <400> 620 cattcacctc ccttagaagc 20 <210> 621 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 621 aacctttaaa tgggtagag 19 <210> 622 <211> 24 <212> DNA <213> STM common primer <400> 622 gcatgccctg cccctaagaa ttcg 24 <210> 623 <211> 20 <212> DNA <213> CNAG_07797 5 flanking region primer 1 <400> 623 agggagaaca ccacaggatg 20 <210> 624 <211> 38 <212> DNA <213> CNAG_07797 5 flanking region primer 2 <400> 624 tcactggccg tcgttttaca ggaatggagg tggtgaac 38 <210> 625 <211> 40 <212> DNA <213> CNAG_07797 3 flanking region primer 1 <400> 625 catggtcata gctgtttcct gggaaagtca gtgatgctcc 40 <210> 626 <211> 18 <212> DNA <213> CNAG_07797 3 flanking region primer 2 <400> 626 gctttcttcc ttttcccg 18 <210> 627 <211> 18 <212> DNA <213> CNAG_07797 diagnostic screening primer, pairing with B79 <400> 627 atcaatggtt tcgtcggc 18 <210> 628 <211> 18 <212> DNA <213> CNAG_07797 Southern blot probe primer <400> 628 ggggcgttga aaatgatg 18 <210> 629 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 629 gagagatccc aacatcacgc 20 <210> 630 <211> 24 <212> DNA <213> STM common primer <400> 630 gcatgccctg cccctaagaa ttcg 24 <210> 631 <211> 22 <212> DNA <213> CNAG_05538 5 flanking region primer 1 <400> 631 ggtctcaaca aagcacttac tg 22 <210> 632 <211> 40 <212> DNA <213> CNAG_05538 5 flanking region primer 2 <400> 632 tcactggccg tcgttttacg taatagtcag gcggtttctc 40 <210> 633 <211> 41 <212> DNA <213> CNAG_05538 3 flanking region primer 1 <400> 633 catggtcata gctgtttcct gagttaccag aggaggagtg g 41 <210> 634 <211> 20 <212> DNA <213> CNAG_05538 3 flanking region primer 2 <400> 634 gattctcttt accctcagcc 20 <210> 635 <211> 20 <212> DNA <213> CNAG_05538 diagnostic screening primer, pairing with B79 <400> 635 tccgattcta ttgactctgc 20 <210> 636 <211> 20 <212> DNA <213> CNAG_05538 Southern blot probe primer <400> 636 gtatcatcca ttttcctggc 20 <210> 637 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 637 ggtgttggat cggggtggat 20 <210> 638 <211> 24 <212> DNA <213> STM common primer <400> 638 gcatgccctg cccctaagaa ttcg 24 <210> 639 <211> 18 <212> DNA <213> CNAG_00332 5 flanking region primer 1 <400> 639 tgattgtcgt ccttgtgg 18 <210> 640 <211> 39 <212> DNA <213> CNAG_00332 5 flanking region primer 2 <400> 640 gctcactggc cgtcgtttta ctttgcgtga ctgttctgc 39 <210> 641 <211> 42 <212> DNA <213> CNAG_00332 3 flanking region primer 1 <400> 641 catggtcata gctgtttcct gcggtttgcg aagatacaga ac 42 <210> 642 <211> 18 <212> DNA <213> CNAG_00332 3 flanking region primer 2 <400> 642 tatggatgcg tgggaatg 18 <210> 643 <211> 18 <212> DNA <213> CNAG_00332 diagnostic screening primer, pairing with B79 <400> 643 gccagaccaa tcaaatgc 18 <210> 644 <211> 19 <212> DNA <213> CNAG_00332 Southern blot probe primer <400> 644 gaagaagggt tgaatgtcg 19 <210> 645 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 645 accgacagct cgaacaagca agag 24 <210> 646 <211> 24 <212> DNA <213> STM common primer <400> 646 gcatgccctg cccctaagaa ttcg 24 <210> 647 <211> 19 <212> DNA <213> CNAG_04023 5 flanking region primer 1 <400> 647 agcaggatag acagcgaag 19 <210> 648 <211> 35 <212> DNA <213> CNAG_04023 5 flanking region primer 2 <400> 648 ctggccgtcg ttttactgtc cttgataggt cggag 35 <210> 649 <211> 35 <212> DNA <213> CNAG_04023 3 flanking region primer 1 <400> 649 gtcatagctg tttcctgtgg aacacattgg aggag 35 <210> 650 <211> 18 <212> DNA <213> CNAG_04023 3 flanking region primer 2 <400> 650 aatgggtgta tgatggcg 18 <210> 651 <211> 19 <212> DNA <213> CNAG_04023 diagnostic screening primer, pairing with B79 <400> 651 cagaacgcat tgtttgtgc 19 <210> 652 <211> 19 <212> DNA <213> CNAG_04023 Southern blot probe primer <400> 652 gtagtaaacg ccttgagcc 19 <210> 653 <211> 20 <212> DNA <213> CNAG_05311 5 flanking region primer 1 <400> 653 caacaagttc catcacatcg 20 <210> 654 <211> 40 <212> DNA <213> CNAG_05311 5 flanking region primer 2 <400> 654 gctcactggc cgtcgtttta cgggtaaaac ctggcttagg 40 <210> 655 <211> 39 <212> DNA <213> CNAG_05311 3 flanking region primer 1 <400> 655 catggtcata gctgtttcct gggatttgga tgaacctgg 39 <210> 656 <211> 21 <212> DNA <213> CNAG_05311 3 flanking region primer 2 <400> 656 gagaaagagg tttgtcgtga g 21 <210> 657 <211> 18 <212> DNA <213> CNAG_05311 diagnostic screening primer, pairing with B79 <400> 657 ttctactttc cgccaagg 18 <210> 658 <211> 19 <212> DNA <213> CNAG_05311 Southern blot probe primer <400> 658 gagatgggtc actaatcgc 19 <210> 659 <211> 19 <212> DNA <213> CNAG_03086 5 flanking region primer 1 <400> 659 aatctccttt gcccgactc 19 <210> 660 <211> 38 <212> DNA <213> CNAG_03086 5 flanking region primer 2 <400> 660 tcactggccg tcgttttaca taccttgtct ttgcgggc 38 <210> 661 <211> 41 <212> DNA <213> CNAG_03086 3 flanking region primer 1 <400> 661 catggtcata gctgtttcct gattgaaggt ggcagtagat g 41 <210> 662 <211> 18 <212> DNA <213> CNAG_03086 3 flanking region primer 2 <400> 662 aaaatgtggg cggatgag 18 <210> 663 <211> 19 <212> DNA <213> CNAG_03086 diagnostic screening primer, pairing with B79 <400> 663 tcagtcggga agagtttgc 19 <210> 664 <211> 19 <212> DNA <213> CNAG_03086 Southern blot probe primer <400> 664 aggcgagagt aagaatgcg 19 <210> 665 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 665 atatggatgt ttttagcgag 20 <210> 666 <211> 24 <212> DNA <213> STM common primer <400> 666 gcatgccctg cccctaagaa ttcg 24 <210> 667 <211> 18 <212> DNA <213> CNAG_03336 5 flanking region primer 1 <400> 667 gtgccttatt ccaatggg 18 <210> 668 <211> 34 <212> DNA <213> CNAG_03336 5 flanking region primer 2 <400> 668 ctggccgtcg ttttacttct tgctgcctgt ttgg 34 <210> 669 <211> 35 <212> DNA <213> CNAG_03336 3 flanking region primer 1 <400> 669 ctggccgtcg ttttacccaa caataatcac gcagg 35 <210> 670 <211> 20 <212> DNA <213> CNAG_03336 3 flanking region primer 2 <400> 670 ccatcctttt tttagcggtc 20 <210> 671 <211> 19 <212> DNA <213> CNAG_03336 diagnostic screening primer, pairing with B79 <400> 671 cgaggtgatt gatgtgagc 19 <210> 672 <211> 20 <212> DNA <213> CNAG_03336 Southern blot probe primer <400> 672 ttcaagagcg gactggacag 20 <210> 673 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 673 gatctctcgc gcttggggga 20 <210> 674 <211> 24 <212> DNA <213> STM common primer <400> 674 gcatgccctg cccctaagaa ttcg 24 <210> 675 <211> 19 <212> DNA <213> CNAG_03229 5 flanking region primer 1 <400> 675 gtgggttact ttgggtgag 19 <210> 676 <211> 37 <212> DNA <213> CNAG_03229 5 flanking region primer 2 <400> 676 tcactggccg tcgttttaca aaagggactt gctgtcg 37 <210> 677 <211> 40 <212> DNA <213> CNAG_03229 3 flanking region primer 1 <400> 677 catggtcata gctgtttcct ggaaagaaag gaaatcgccg 40 <210> 678 <211> 18 <212> DNA <213> CNAG_03229 3 flanking region primer 2 <400> 678 tggaagatgg tgacgaag 18 <210> 679 <211> 18 <212> DNA <213> CNAG_03229 diagnostic screening primer, pairing with B79 <400> 679 ttgcgatttc cgactacc 18 <210> 680 <211> 18 <212> DNA <213> CNAG_03229 Southern blot probe primer <400> 680 tggaggcgaa taatggtc 18 <210> 681 <211> 20 <212> DNA <213> NAT#227 STM primer <400> 681 tcgtggttta gagggagcgc 20 <210> 682 <211> 24 <212> DNA <213> STM common primer <400> 682 gcatgccctg cccctaagaa ttcg 24 <210> 683 <211> 19 <212> DNA <213> CNAG_01883 5 flanking region primer 1 <400> 683 tactcgctgc ttagggttc 19 <210> 684 <211> 35 <212> DNA <213> CNAG_01883 5 flanking region primer 2 <400> 684 ctggccgtcg ttttacatgg catactccag gtctg 35 <210> 685 <211> 36 <212> DNA <213> CNAG_01883 3 flanking region primer 1 <400> 685 gtcatagctg tttcctgccc tccatttact tctccg 36 <210> 686 <211> 22 <212> DNA <213> CNAG_01883 3 flanking region primer 2 <400> 686 tagcagtagg gctcctgatt cg 22 <210> 687 <211> 19 <212> DNA <213> CNAG_01883 diagnostic screening primer, pairing with B79 <400> 687 ttggaggtct ttatcagcg 19 <210> 688 <211> 19 <212> DNA <213> CNAG_01883 Southern blot probe primer <400> 688 ggagaagtaa atggagggg 19 <210> 689 <211> 24 <212> DNA <213> NAT#125 STM primer <400> 689 cgctacagcc agcgcgcgca agcg 24 <210> 690 <211> 24 <212> DNA <213> STM common primer <400> 690 gcatgccctg cccctaagaa ttcg 24 <210> 691 <211> 20 <212> DNA <213> CNAG_04837 5 flanking region primer 1 <400> 691 tgaaacagtg aatcaggcac 20 <210> 692 <211> 40 <212> DNA <213> CNAG_04837 5 flanking region primer 2 <400> 692 gctcactggc cgtcgtttta ctcagtgtca acggtcattg 40 <210> 693 <211> 42 <212> DNA <213> CNAG_04837 3 flanking region primer 1 <400> 693 catggtcata gctgtttcct ggaaaggggt agggataagt tg 42 <210> 694 <211> 20 <212> DNA <213> CNAG_04837 3 flanking region primer 2 <400> 694 actgaacgac aagagacgac 20 <210> 695 <211> 18 <212> DNA <213> CNAG_04837 diagnostic screening primer, pairing with B79 <400> 695 taagagaaaa ggggctgc 18 <210> 696 <211> 21 <212> DNA <213> CNAG_04837 Southern blot probe primer 1 <400> 696 tgtccccatc aactctacaa c 21 <210> 697 <211> 20 <212> DNA <213> CNAG_04837 Southern blot probe primer 2 <400> 697 cttctgcttt caccattagc 20 <210> 698 <211> 21 <212> DNA <213> NAT#146 STM primer <400> 698 actagccccc cctcaccacc t 21 <210> 699 <211> 24 <212> DNA <213> STM common primer <400> 699 gcatgccctg cccctaagaa ttcg 24 <210> 700 <211> 19 <212> DNA <213> CNAG_00514 5 flanking region primer 1 <400> 700 ggtgtttcca tacgaaccg 19 <210> 701 <211> 38 <212> DNA <213> CNAG_00514 5 flanking region primer 2 <400> 701 ctggccgtcg ttttacggag ccatttctta cttatgcg 38 <210> 702 <211> 35 <212> DNA <213> CNAG_00514 3 flanking region primer 1 <400> 702 gtcatagctg tttcctgaat cccaaggacc atctg 35 <210> 703 <211> 20 <212> DNA <213> CNAG_00514 3 flanking region primer 2 <400> 703 gctgagaagg agaggatagc 20 <210> 704 <211> 20 <212> DNA <213> CNAG_00514 diagnostic screening primer, pairing with B79 <400> 704 gtggttgaaa gtattaggcg 20 <210> 705 <211> 19 <212> DNA <213> CNAG_00514 Southern blot probe primer <400> 705 gaggattggt gatggactg 19 <210> 706 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 706 caccctctat ctcgagaaag ctcc 24 <210> 707 <211> 24 <212> DNA <213> STM common primer <400> 707 gcatgccctg cccctaagaa ttcg 24 <210> 708 <211> 20 <212> DNA <213> CNAG_03409 5 flanking region primer 1 <400> 708 cctcaccgat tcattcactg 20 <210> 709 <211> 38 <212> DNA <213> CNAG_03409 5 flanking region primer 2 <400> 709 ctggccgtcg ttttactttt ctcccttctt cgcccgac 38 <210> 710 <211> 37 <212> DNA <213> CNAG_03409 3 flanking region primer 1 <400> 710 gtcatagctg tttcctgagg ttcaagccca gactcag 37 <210> 711 <211> 20 <212> DNA <213> CNAG_03409 3 flanking region primer 2 <400> 711 agtatctacg gtgctggtgc 20 <210> 712 <211> 20 <212> DNA <213> CNAG_03409 diagnostic screening primer, pairing with B79 <400> 712 tgaggatgat gactggggac 20 <210> 713 <211> 20 <212> DNA <213> CNAG_03409 Southern blot probe primer 1 <400> 713 acttccaatg ctcaaccccg 20 <210> 714 <211> 20 <212> DNA <213> CNAG_03409 Southern blot probe primer 2 <400> 714 gcaggacaag aaggagagtg 20 <210> 715 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 715 caccctctat ctcgagaaag ctcc 24 <210> 716 <211> 24 <212> DNA <213> STM common primer <400> 716 gcatgccctg cccctaagaa ttcg 24 <210> 717 <211> 19 <212> DNA <213> CNAG_05392 5 flanking region primer 1 <400> 717 aagttgtgaa accaccagg 19 <210> 718 <211> 38 <212> DNA <213> CNAG_05392 5 flanking region primer 2 <400> 718 tcactggccg tcgttttact caccctgtag atgtgtgg 38 <210> 719 <211> 42 <212> DNA <213> CNAG_05392 3 flanking region primer 1 <400> 719 catggtcata gctgtttcct gggagtccct gtgaagtaaa ag 42 <210> 720 <211> 20 <212> DNA <213> CNAG_05392 3 flanking region primer 2 <400> 720 tgagccaaag tagcagtcag 20 <210> 721 <211> 19 <212> DNA <213> CNAG_05392 diagnostic screening primer, pairing with B79 <400> 721 gaatggacct tggaaaacg 19 <210> 722 <211> 20 <212> DNA <213> CNAG_05392 Southern blot probe primer <400> 722 cggtctgtgt gcttcttcac 20 <210> 723 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 723 gatctctcgc gcttggggga 20 <210> 724 <211> 24 <212> DNA <213> STM common primer <400> 724 gcatgccctg cccctaagaa ttcg 24 <210> 725 <211> 20 <212> DNA <213> CNAG_04895 5 flanking region primer 1 <400> 725 ttcgtagtta gccagcagag 20 <210> 726 <211> 38 <212> DNA <213> CNAG_04895 5 flanking region primer 2 <400> 726 tcactggccg tcgttttacg ttcttgatgt ggaggagc 38 <210> 727 <211> 40 <212> DNA <213> CNAG_04895 3 flanking region primer 1 <400> 727 catggtcata gctgtttcct gcccaaccct tatgcgtatg 40 <210> 728 <211> 18 <212> DNA <213> CNAG_04895 3 flanking region primer 2 <400> 728 agcgtctccc tcaaaaac 18 <210> 729 <211> 18 <212> DNA <213> CNAG_04895 diagnostic screening primer, pairing with B79 <400> 729 ttagcgaccg aaaaaggg 18 <210> 730 <211> 19 <212> DNA <213> CNAG_04895 Southern blot probe primer <400> 730 aagatgacgc agaactggg 19 <210> 731 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 731 gatctctcgc gcttggggga 20 <210> 732 <211> 24 <212> DNA <213> STM common primer <400> 732 gcatgccctg cccctaagaa ttcg 24 <210> 733 <211> 19 <212> DNA <213> CNAG_04176 5 flanking region primer 1 <400> 733 ttgacacggg gactaatcg 19 <210> 734 <211> 40 <212> DNA <213> CNAG_04176 5 flanking region primer 2 <400> 734 gctcactggc cgtcgtttta cgattagccc agtcttagcg 40 <210> 735 <211> 41 <212> DNA <213> CNAG_04176 3 flanking region primer 1 <400> 735 catggtcata gctgtttcct gagaagcagg gagcaaaacg g 41 <210> 736 <211> 19 <212> DNA <213> CNAG_04176 3 flanking region primer 2 <400> 736 ccgaaaagtt gagcaagac 19 <210> 737 <211> 23 <212> DNA <213> CNAG_04176 diagnostic screening primer, pairing with B79 <400> 737 cgatagcctt tgtagtaggt ctc 23 <210> 738 <211> 20 <212> DNA <213> CNAG_04176 Southern blot probe primer <400> 738 gggatttgat tatctggagg 20 <210> 739 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 739 tatccccctc tccgctctct agca 24 <210> 740 <211> 24 <212> DNA <213> STM common primer <400> 740 gcatgccctg cccctaagaa ttcg 24 <210> 741 <211> 19 <212> DNA <213> CNAG_07724 5 flanking region primer 1 <400> 741 caaagaaaac gctcactcc 19 <210> 742 <211> 40 <212> DNA <213> CNAG_07724 5 flanking region primer 2 <400> 742 tcactggccg tcgttttacg gatagaacaa gacaagagcg 40 <210> 743 <211> 41 <212> DNA <213> CNAG_07724 3 flanking region primer 1 <400> 743 catggtcata gctgtttcct gacttcagtc gttaccagca c 41 <210> 744 <211> 19 <212> DNA <213> CNAG_07724 3 flanking region primer 2 <400> 744 tctttgagca catcaggtg 19 <210> 745 <211> 19 <212> DNA <213> CNAG_07724 diagnostic screening primer, pairing with B79 <400> 745 ccgtcattat cgtgtctcc 19 <210> 746 <211> 19 <212> DNA <213> CNAG_07724 Southern blot probe primer <400> 746 gcttcggagg agttgatag 19 <210> 747 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 747 tatccccctc tccgctctct agca 24 <210> 748 <211> 24 <212> DNA <213> STM common primer <400> 748 gcatgccctg cccctaagaa ttcg 24 <210> 749 <211> 19 <212> DNA <213> CNAG_06751 5 flanking region primer 1 <400> 749 aaaaacggct gaaggctcg 19 <210> 750 <211> 40 <212> DNA <213> CNAG_06751 5 flanking region primer 2 <400> 750 gctcactggc cgtcgtttta ctggcgagtt ggtatctgag 40 <210> 751 <211> 40 <212> DNA <213> CNAG_06751 3 flanking region primer 1 <400> 751 catggtcata gctgtttcct gaagatgaga gagaaggggg 40 <210> 752 <211> 20 <212> DNA <213> CNAG_06751 3 flanking region primer 2 <400> 752 agaagtggta tgaccgtttg 20 <210> 753 <211> 19 <212> DNA <213> CNAG_06751 diagnostic screening primer, pairing with B79 <400> 753 tccatacatt gttgagggc 19 <210> 754 <211> 18 <212> DNA <213> CNAG_06751 Southern blot probe primer <400> 754 attgcccaag cggataag 18 <210> 755 <211> 22 <212> DNA <213> NAT#208 STM primer <400> 755 tggtcgcggg agatcgtggt tt 22 <210> 756 <211> 24 <212> DNA <213> STM common primer <400> 756 gcatgccctg cccctaagaa ttcg 24 <210> 757 <211> 18 <212> DNA <213> CNAG_01438 5 flanking region primer 1 <400> 757 atcacacgca atcccatc 18 <210> 758 <211> 39 <212> DNA <213> CNAG_01438 5 flanking region primer 2 <400> 758 gctcactggc cgtcgtttta cagcaagatg gaaggaagc 39 <210> 759 <211> 39 <212> DNA <213> CNAG_01438 3 flanking region primer 1 <400> 759 catggtcata gctgtttcct ggcgacgaaa tcataaggc 39 <210> 760 <211> 19 <212> DNA <213> CNAG_01438 3 flanking region primer 2 <400> 760 tgagaactgg acgaactgc 19 <210> 761 <211> 18 <212> DNA <213> CNAG_01438 diagnostic screening primer, pairing with B79 <400> 761 aatgatgcgt cggacttc 18 <210> 762 <211> 20 <212> DNA <213> CNAG_01438 Southern blot probe primer <400> 762 ggaagagaaa gacttgcgag 20 <210> 763 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 763 acagctccaa acctcgctaa acag 24 <210> 764 <211> 24 <212> DNA <213> STM common primer <400> 764 gcatgccctg cccctaagaa ttcg 24 <210> 765 <211> 20 <212> DNA <213> CNAG_00791 5 flanking region primer 1 <400> 765 gcatccattt tatcggttcg 20 <210> 766 <211> 41 <212> DNA <213> CNAG_00791 5 flanking region primer 2 <400> 766 gctcactggc cgtcgtttta cttctgcttg aaggagtcgg g 41 <210> 767 <211> 40 <212> DNA <213> CNAG_00791 3 flanking region primer 1 <400> 767 catggtcata gctgtttcct gcggttatct tgcgtaaagc 40 <210> 768 <211> 19 <212> DNA <213> CNAG_00791 3 flanking region primer 2 <400> 768 tgtctcaccc atcacgaac 19 <210> 769 <211> 18 <212> DNA <213> CNAG_00791 diagnostic screening primer, pairing with B79 <400> 769 attcgctcag gcagtttc 18 <210> 770 <211> 19 <212> DNA <213> CNAG_00791 Southern blot probe primer <400> 770 gcgatgatga ggactcttg 19 <210> 771 <211> 21 <212> DNA <213> NAT#146 STM primer <400> 771 actagccccc cctcaccacc t 21 <210> 772 <211> 24 <212> DNA <213> STM common primer <400> 772 gcatgccctg cccctaagaa ttcg 24 <210> 773 <211> 19 <212> DNA <213> CNAG_02241 5 flanking region primer 1 <400> 773 atagcaaacc catacctcg 19 <210> 774 <211> 37 <212> DNA <213> CNAG_02241 5 flanking region primer 2 <400> 774 tcactggccg tcgttttact ttgtttggta gtggggg 37 <210> 775 <211> 40 <212> DNA <213> CNAG_02241 3 flanking region primer 1 <400> 775 catggtcata gctgtttcct gcgagaactc tggagcattc 40 <210> 776 <211> 21 <212> DNA <213> CNAG_02241 3 flanking region primer 2 <400> 776 gatgtgttgt ggactgtatg c 21 <210> 777 <211> 20 <212> DNA <213> CNAG_02241 diagnostic screening primer, pairing with B79 <400> 777 tcccatcata ggagtggacg 20 <210> 778 <211> 20 <212> DNA <213> CNAG_02241 Southern blot probe primer <400> 778 cttgggagca aataagtgtg 20 <210> 779 <211> 20 <212> DNA <213> NAT#219 STM primer <400> 779 ccctaaaacc ctacagcaat 20 <210> 780 <211> 24 <212> DNA <213> STM common primer <400> 780 gcatgccctg cccctaagaa ttcg 24 <210> 781 <211> 19 <212> DNA <213> CNAG_02566 5 flanking region primer 1 <400> 781 tcgcacggca taggattag 19 <210> 782 <211> 38 <212> DNA <213> CNAG_02566 5 flanking region primer 2 <400> 782 tcactggccg tcgttttaca tgccgccaga atgttgag 38 <210> 783 <211> 40 <212> DNA <213> CNAG_02566 3 flanking region primer 1 <400> 783 catggtcata gctgtttcct gtgatgttga tggagactgg 40 <210> 784 <211> 20 <212> DNA <213> CNAG_02566 3 flanking region primer 2 <400> 784 actgcttctt caccctccac 20 <210> 785 <211> 18 <212> DNA <213> CNAG_02566 diagnostic screening primer, pairing with B79 <400> 785 atctgccgca acattcac 18 <210> 786 <211> 21 <212> DNA <213> CNAG_02566 Southern blot probe primer <400> 786 gttggtgctt catagtgact g 21 <210> 787 <211> 20 <212> DNA <213> NAT#219 STM primer <400> 787 ccctaaaacc ctacagcaat 20 <210> 788 <211> 24 <212> DNA <213> STM common primer <400> 788 gcatgccctg cccctaagaa ttcg 24 <210> 789 <211> 19 <212> DNA <213> CNAG_01626 5 flanking region primer 1 <400> 789 ggatgatgga atcgtatgc 19 <210> 790 <211> 38 <212> DNA <213> CNAG_01626 5 flanking region primer 2 <400> 790 tcactggccg tcgttttaca ttatccaccc tcggcttc 38 <210> 791 <211> 42 <212> DNA <213> CNAG_01626 3 flanking region primer 1 <400> 791 catggtcata gctgtttcct ggcgaacggt atgacaacta tg 42 <210> 792 <211> 22 <212> DNA <213> CNAG_01626 3 flanking region primer 2 <400> 792 ggacgaaaac attgctctca ac 22 <210> 793 <211> 19 <212> DNA <213> CNAG_01626 diagnostic screening primer, pairing with B79 <400> 793 tcagcagcag caggtaaac 19 <210> 794 <211> 18 <212> DNA <213> CNAG_01626 Southern blot probe primer <400> 794 cccagaccgt tttgaatg 18 <210> 795 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 795 ctttaaaggt ggtttgtg 18 <210> 796 <211> 24 <212> DNA <213> STM common primer <400> 796 gcatgccctg cccctaagaa ttcg 24 <210> 797 <211> 20 <212> DNA <213> CNAG_05940 5 flanking region primer 1 <400> 797 tcttgaccgt cgccagttac 20 <210> 798 <211> 39 <212> DNA <213> CNAG_05940 5 flanking region primer 2 <400> 798 tcactggccg tcgttttact ggctcaaagg caaagtcgg 39 <210> 799 <211> 40 <212> DNA <213> CNAG_05940 3 flanking region primer 1 <400> 799 catggtcata gctgtttcct gaggtgattt tgggagtcag 40 <210> 800 <211> 19 <212> DNA <213> CNAG_05940 3 flanking region primer 2 <400> 800 ggctgagaat gatgtgacg 19 <210> 801 <211> 19 <212> DNA <213> CNAG_05940 diagnostic screening primer, pairing with B79 <400> 801 ggaaagaaga cggatagcg 19 <210> 802 <211> 19 <212> DNA <213> CNAG_05940 Southern blot probe primer <400> 802 gtctccagga aagttcgtg 19 <210> 803 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 803 ctttaaaggt ggtttgtg 18 <210> 804 <211> 24 <212> DNA <213> STM common primer <400> 804 gcatgccctg cccctaagaa ttcg 24 <210> 805 <211> 22 <212> DNA <213> CNAG_04352 5 flanking region primer 1 <400> 805 gtgattatgt gaacccgtta cc 22 <210> 806 <211> 39 <212> DNA <213> CNAG_04352 5 flanking region primer 2 <400> 806 tcactggccg tcgttttact tttagaggtg atgactgcg 39 <210> 807 <211> 41 <212> DNA <213> CNAG_04352 3 flanking region primer 1 <400> 807 catggtcata gctgtttcct gtgctgctgg aggaagagaa c 41 <210> 808 <211> 19 <212> DNA <213> CNAG_04352 3 flanking region primer 2 <400> 808 agtgagtcgt tgatgtggc 19 <210> 809 <211> 19 <212> DNA <213> CNAG_04352 diagnostic screening primer, pairing with B79 <400> 809 atagtgaata cgggtcggg 19 <210> 810 <211> 19 <212> DNA <213> CNAG_04352 Southern blot probe primer <400> 810 gctgaaacag gaggcatag 19 <210> 811 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 811 ccagccagca ctccgatata 20 <210> 812 <211> 24 <212> DNA <213> STM common primer <400> 812 gcatgccctg cccctaagaa ttcg 24 <210> 813 <211> 18 <212> DNA <213> CNAG_03768 5 flanking region primer 1 <400> 813 aatgtccttg cggttgac 18 <210> 814 <211> 40 <212> DNA <213> CNAG_03768 5 flanking region primer 2 <400> 814 gctcactggc cgtcgtttta caatagtccc cgtcgttgtc 40 <210> 815 <211> 41 <212> DNA <213> CNAG_03768 3 flanking region primer 1 <400> 815 catggtcata gctgtttcct gcgatggatg tttggtggta g 41 <210> 816 <211> 19 <212> DNA <213> CNAG_03768 3 flanking region primer 2 <400> 816 aaaggggttt tggtggctc 19 <210> 817 <211> 20 <212> DNA <213> CNAG_03768 diagnostic screening primer, pairing with B79 <400> 817 tggatttggc agagaagaag 20 <210> 818 <211> 19 <212> DNA <213> CNAG_03768 Southern blot probe primer <400> 818 aagacacttg gctctccag 19 <210> 819 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 819 ccagccagca ctccgatata 20 <210> 820 <211> 24 <212> DNA <213> STM common primer <400> 820 gcatgccctg cccctaagaa ttcg 24 <210> 821 <211> 20 <212> DNA <213> CNAG_02322 5 flanking region primer 1 <400> 821 cgctcactca cctttcaaac 20 <210> 822 <211> 35 <212> DNA <213> CNAG_02322 5 flanking region primer 2 <400> 822 ctggccgtcg ttttacacat cggcttttac ggagc 35 <210> 823 <211> 37 <212> DNA <213> CNAG_02322 3 flanking region primer 1 <400> 823 gtcatagctg tttcctgctc ggttaccagt cactacg 37 <210> 824 <211> 20 <212> DNA <213> CNAG_02322 3 flanking region primer 2 <400> 824 ccaccaagat tcagagttcg 20 <210> 825 <211> 19 <212> DNA <213> CNAG_02322 diagnostic screening primer, pairing with B79 <400> 825 gagatttcgg tggatttcc 19 <210> 826 <211> 18 <212> DNA <213> CNAG_02322 Southern blot probe primer <400> 826 aagcaaccct cagcattg 18 <210> 827 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 827 ggtgttggat cggggtggat 20 <210> 828 <211> 24 <212> DNA <213> STM common primer <400> 828 gcatgccctg cccctaagaa ttcg 24 <210> 829 <211> 20 <212> DNA <213> CNAG_06818 5 flanking region primer 1 <400> 829 tgtattctgc ccacctccac 20 <210> 830 <211> 39 <212> DNA <213> CNAG_06818 5 flanking region primer 2 <400> 830 tcactggccg tcgttttacc ctctttgctg gtctctgag 39 <210> 831 <211> 41 <212> DNA <213> CNAG_06818 3 flanking region primer 1 <400> 831 catggtcata gctgtttcct gtctgctgct acatcaggtt c 41 <210> 832 <211> 19 <212> DNA <213> CNAG_06818 3 flanking region primer 2 <400> 832 aacctctgta tcgctgacg 19 <210> 833 <211> 22 <212> DNA <213> CNAG_06818 diagnostic screening primer, pairing with B79 <400> 833 gggtgaaaga caataggtaa gg 22 <210> 834 <211> 21 <212> DNA <213> CNAG_06818 Southern blot probe primer <400> 834 aaagtgaaga gaggaggtca g 21 <210> 835 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 835 ggtgttggat cggggtggat 20 <210> 836 <211> 24 <212> DNA <213> STM common primer <400> 836 gcatgccctg cccctaagaa ttcg 24 <210> 837 <211> 19 <212> DNA <213> CNAG_07011 5 flanking region primer 1 <400> 837 ccaacaaata gcaaagccg 19 <210> 838 <211> 38 <212> DNA <213> CNAG_07011 5 flanking region primer 2 <400> 838 tcactggccg tcgttttaca caccaaagcc aagaacac 38 <210> 839 <211> 41 <212> DNA <213> CNAG_07011 3 flanking region primer 1 <400> 839 catggtcata gctgtttcct ggctaatgcc tgaaacagac c 41 <210> 840 <211> 19 <212> DNA <213> CNAG_07011 3 flanking region primer 2 <400> 840 tcggacaccc aatcaatac 19 <210> 841 <211> 18 <212> DNA <213> CNAG_07011 diagnostic screening primer, pairing with B79 <400> 841 ttgtgttgtt cggtgagg 18 <210> 842 <211> 20 <212> DNA <213> CNAG_07011 Southern blot probe primer <400> 842 tgggtgatac agaggaaaac 20 <210> 843 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 843 gagatctttc gggaggtctg gatt 24 <210> 844 <211> 24 <212> DNA <213> STM common primer <400> 844 gcatgccctg cccctaagaa ttcg 24 <210> 845 <211> 20 <212> DNA <213> CNAG_04036 5 flanking region primer 1 <400> 845 ccagttgaac gcttctaaac 20 <210> 846 <211> 39 <212> DNA <213> CNAG_04036 5 flanking region primer 2 <400> 846 tcactggccg tcgttttact gatggtgtgg gaacaatac 39 <210> 847 <211> 40 <212> DNA <213> CNAG_04036 3 flanking region primer 1 <400> 847 catggtcata gctgtttcct gttctccttg cctatgtgcc 40 <210> 848 <211> 20 <212> DNA <213> CNAG_04036 3 flanking region primer 2 <400> 848 cagcaactta ctttggaggc 20 <210> 849 <211> 19 <212> DNA <213> CNAG_04036 diagnostic screening primer, pairing with B79 <400> 849 atagacagca gcgggatac 19 <210> 850 <211> 18 <212> DNA <213> CNAG_04036 Southern blot probe primer <400> 850 tcacagatgc ctttctcg 18 <210> 851 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 851 gagatctttc gggaggtctg gatt 24 <210> 852 <211> 24 <212> DNA <213> STM common primer <400> 852 gcatgccctg cccctaagaa ttcg 24 <210> 853 <211> 19 <212> DNA <213> CNAG_07506 5 flanking region primer 1 <400> 853 cgtaatgaca ctggagagc 19 <210> 854 <211> 42 <212> DNA <213> CNAG_07506 5 flanking region primer 2 <400> 854 gctcactggc cgtcgtttta ctgttgccgt aggaaccgct tc 42 <210> 855 <211> 40 <212> DNA <213> CNAG_07506 3 flanking region primer 1 <400> 855 catggtcata gctgtttcct gtcccaaagc ctcttctcag 40 <210> 856 <211> 18 <212> DNA <213> CNAG_07506 3 flanking region primer 2 <400> 856 gtgagaccct gatagacc 18 <210> 857 <211> 19 <212> DNA <213> CNAG_07506 diagnostic screening primer, pairing with B79 <400> 857 gactcaaagg gagatgacg 19 <210> 858 <211> 19 <212> DNA <213> CNAG_07506 Southern blot probe primer <400> 858 gcttttgcct gctattgac 19 <210> 859 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 859 cgcccgccct cactatccac 20 <210> 860 <211> 24 <212> DNA <213> STM common primer <400> 860 gcatgccctg cccctaagaa ttcg 24 <210> 861 <211> 19 <212> DNA <213> CNAG_06223 5 flanking region primer 1 <400> 861 ggatgaaggg gaagacttg 19 <210> 862 <211> 38 <212> DNA <213> CNAG_06223 5 flanking region primer 2 <400> 862 tcactggccg tcgttttacc ttccgttcaa aatcgtcg 38 <210> 863 <211> 41 <212> DNA <213> CNAG_06223 3 flanking region primer 1 <400> 863 catggtcata gctgtttcct ggtctaccac caccatcaca c 41 <210> 864 <211> 20 <212> DNA <213> CNAG_06223 3 flanking region primer 2 <400> 864 gccgtccatt catacatctg 20 <210> 865 <211> 19 <212> DNA <213> CNAG_06223 diagnostic screening primer, pairing with B79 <400> 865 ggtgtcaata gttgggtgg 19 <210> 866 <211> 19 <212> DNA <213> CNAG_06223 Southern blot probe primer <400> 866 agagcagaag caaatgacc 19 <210> 867 <211> 20 <212> DNA <213> NAT#210 STM primer <400> 867 ctagagcccg ccacaacgct 20 <210> 868 <211> 24 <212> DNA <213> STM common primer <400> 868 gcatgccctg cccctaagaa ttcg 24 <210> 869 <211> 24 <212> DNA <213> CNAG_06283 5 flanking region primer 1 <400> 869 ccctttgtcc ttacccacat tttc 24 <210> 870 <211> 39 <212> DNA <213> CNAG_06283 5 flanking region primer 2 <400> 870 tcactggccg tcgttttact cgttcgggca cttcttgac 39 <210> 871 <211> 41 <212> DNA <213> CNAG_06283 3 flanking region primer 1 <400> 871 catggtcata gctgtttcct gcgagtaagg agacggttga g 41 <210> 872 <211> 22 <212> DNA <213> CNAG_06283 3 flanking region primer 2 <400> 872 ggttaggggg ataaaatagg ag 22 <210> 873 <211> 21 <212> DNA <213> CNAG_06283 diagnostic screening primer, pairing with B79 <400> 873 gagacattct ttctgacgag c 21 <210> 874 <211> 20 <212> DNA <213> CNAG_06283 Southern blot probe primer <400> 874 tggcttgaag ggtcttggtc 20 <210> 875 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 875 ccagccagca ctccgatata 20 <210> 876 <211> 24 <212> DNA <213> STM common primer <400> 876 gcatgccctg cccctaagaa ttcg 24 <210> 877 <211> 22 <212> DNA <213> CNAG_04583 5 flanking region primer 1 <400> 877 gctcgctaca ttttcaggta tc 22 <210> 878 <211> 39 <212> DNA <213> CNAG_04583 5 flanking region primer 2 <400> 878 tcactggccg tcgttttacg agagggtcgg aaatagaag 39 <210> 879 <211> 41 <212> DNA <213> CNAG_04583 3 flanking region primer 1 <400> 879 catggtcata gctgtttcct ggggaaggag taatgcctga c 41 <210> 880 <211> 18 <212> DNA <213> CNAG_04583 3 flanking region primer 2 <400> 880 aaccgtgatg gagattgg 18 <210> 881 <211> 20 <212> DNA <213> CNAG_04583 diagnostic screening primer, pairing with B79 <400> 881 cggtgtgcga ttctaacttc 20 <210> 882 <211> 19 <212> DNA <213> CNAG_04583 Southern blot probe primer <400> 882 ggggactgtt ttgatgacc 19 <210> 883 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 883 ccatagcgat atctacccca atct 24 <210> 884 <211> 24 <212> DNA <213> STM common primer <400> 884 gcatgccctg cccctaagaa ttcg 24 <210> 885 <211> 19 <212> DNA <213> CNAG_03261 5 flanking region primer 1 <400> 885 ggcgataaag acgacgaac 19 <210> 886 <211> 35 <212> DNA <213> CNAG_03261 5 flanking region primer 2 <400> 886 ctggccgtcg ttttacggac tattcgtggc ttagc 35 <210> 887 <211> 35 <212> DNA <213> CNAG_03261 3 flanking region primer 1 <400> 887 gtcatagctg tttcctgtga gaccaatgtg ttgcc 35 <210> 888 <211> 24 <212> DNA <213> CNAG_03261 3 flanking region primer 2 <400> 888 cctcaacccc tttctgattt atcc 24 <210> 889 <211> 22 <212> DNA <213> CNAG_03261 diagnostic screening primer, pairing with B79 <400> 889 tagtcggtga tgagagttgc gg 22 <210> 890 <211> 20 <212> DNA <213> CNAG_03261 Southern blot probe primer <400> 890 gcaaatctgc ttctgggatg 20 <210> 891 <211> 19 <212> DNA <213> CNAG_04908 5 flanking region primer 1 <400> 891 gatgacgaat agcacggtc 19 <210> 892 <211> 37 <212> DNA <213> CNAG_04908 5 flanking region primer 2 <400> 892 tcactggccg tcgttttacg gatgggaaga ttgttgg 37 <210> 893 <211> 39 <212> DNA <213> CNAG_04908 3 flanking region primer 1 <400> 893 gatggtcata gctgtttcct gttgcttacg cctttgtcc 39 <210> 894 <211> 18 <212> DNA <213> CNAG_04908 3 flanking region primer 2 <400> 894 atggttgttc acgcacag 18 <210> 895 <211> 18 <212> DNA <213> CNAG_04908 diagnostic screening primer, pairing with B79 <400> 895 aaacaaaagc cgagggag 18 <210> 896 <211> 19 <212> DNA <213> CNAG_04908 Southern blot probe primer <400> 896 aacttcccac cccttcatc 19 <210> 897 <211> 24 <212> DNA <213> NAT#242 STM primer <400> 897 gtagcgatag gggtgtcgct ttag 24 <210> 898 <211> 24 <212> DNA <213> STM common primer <400> 898 gcatgccctg cccctaagaa ttcg 24 <210> 899 <211> 19 <212> DNA <213> CNAG_01317 5 flanking region primer 1 <400> 899 aagcaccaag aagcaagtg 19 <210> 900 <211> 37 <212> DNA <213> CNAG_01317 5 flanking region primer 2 <400> 900 tcactggccg tcgttttaca ttgacacctg ccgattc 37 <210> 901 <211> 40 <212> DNA <213> CNAG_01317 3 flanking region primer 1 <400> 901 catggtcata gctgtttcct gtactatcag cagaagcggc 40 <210> 902 <211> 20 <212> DNA <213> CNAG_01317 3 flanking region primer 2 <400> 902 aatgataggc ggaagtaagg 20 <210> 903 <211> 19 <212> DNA <213> CNAG_01317 diagnostic screening primer, pairing with B79 <400> 903 ccacctcgca gtatcaaac 19 <210> 904 <211> 20 <212> DNA <213> CNAG_01317 Southern blot probe primer <400> 904 agggtcaggg agatgatgtg 20 <210> 905 <211> 19 <212> DNA <213> CNAG_00559 5 flanking region primer 1 <400> 905 ggtgtttcca tacgaaccg 19 <210> 906 <211> 38 <212> DNA <213> CNAG_00559 5 flanking region primer 2 <400> 906 ctggccgtcg ttttacggag ccatttctta cttatgcg 38 <210> 907 <211> 35 <212> DNA <213> CNAG_00559 3 flanking region primer 1 <400> 907 gtcatagctg tttcctgaat cccaaggacc atctg 35 <210> 908 <211> 20 <212> DNA <213> CNAG_00559 3 flanking region primer 2 <400> 908 gctgagaagg agaggatagc 20 <210> 909 <211> 20 <212> DNA <213> CNAG_00559 diagnostic screening primer, pairing with B79 <400> 909 gtggttgaaa gtattaggcg 20 <210> 910 <211> 19 <212> DNA <213> CNAG_00559 Southern blot probe primer <400> 910 tctcttcgtt tgctacacg 19 <210> 911 <211> 24 <212> DNA <213> NAT#242 STM primer <400> 911 gtagcgatag gggtgtcgct ttag 24 <210> 912 <211> 24 <212> DNA <213> STM common primer <400> 912 gcatgccctg cccctaagaa ttcg 24 <210> 913 <211> 18 <212> DNA <213> CNAG_07940 5 flanking region primer 1 <400> 913 caaggtaatg attggggc 18 <210> 914 <211> 38 <212> DNA <213> CNAG_07940 5 flanking region primer 2 <400> 914 tcactggccg tcgttttacg caataatgga aggtctcg 38 <210> 915 <211> 39 <212> DNA <213> CNAG_07940 3 flanking region primer 1 <400> 915 catggtcata gctgtttcct gatcaagaat gacacgccg 39 <210> 916 <211> 19 <212> DNA <213> CNAG_07940 3 flanking region primer 2 <400> 916 aacttgtatg gtggtgccg 19 <210> 917 <211> 19 <212> DNA <213> CNAG_07940 diagnostic screening primer, pairing with B79 <400> 917 tcacttgctc ttctccctg 19 <210> 918 <211> 18 <212> DNA <213> CNAG_07940 Southern blot probe primer <400> 918 gtgacgaatg cgaaactc 18 <210> 919 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 919 atatggatgt ttttagcgag 20 <210> 920 <211> 24 <212> DNA <213> STM common primer <400> 920 gcatgccctg cccctaagaa ttcg 24 <210> 921 <211> 18 <212> DNA <213> CNAG_05535 5 flanking region primer 1 <400> 921 agcaacaacg gcatagtg 18 <210> 922 <211> 39 <212> DNA <213> CNAG_05535 5 flanking region primer 2 <400> 922 tcactggccg tcgttttaca cgacagtgag agtcctgac 39 <210> 923 <211> 43 <212> DNA <213> CNAG_05535 3 flanking region primer 1 <400> 923 catggtcata gctgtttcct gctggagcga ggaaatagta atg 43 <210> 924 <211> 18 <212> DNA <213> CNAG_05535 3 flanking region primer 2 <400> 924 acacaaagcg agtggttg 18 <210> 925 <211> 19 <212> DNA <213> CNAG_05535 diagnostic screening primer, pairing with B79 <400> 925 ggcagtgtca gatgaaagc 19 <210> 926 <211> 19 <212> DNA <213> CNAG_05535 Southern blot probe primer <400> 926 catcggctac atcttcctc 19 <210> 927 <211> 21 <212> DNA <213> CNAG_02516 5 flanking region primer 1 <400> 927 gcatcacacg aacgattcta c 21 <210> 928 <211> 38 <212> DNA <213> CNAG_02516 5 flanking region primer 2 <400> 928 tcactggccg tcgttttaca ttgttggcgg tggaaacg 38 <210> 929 <211> 41 <212> DNA <213> CNAG_02516 3 flanking region primer 1 <400> 929 catggtcata gctgtttcct gaggtttggg agcagtaagg c 41 <210> 930 <211> 19 <212> DNA <213> CNAG_02516 3 flanking region primer 2 <400> 930 gatgtttgtg accgattcg 19 <210> 931 <211> 19 <212> DNA <213> CNAG_02516 diagnostic screening primer, pairing with B79 <400> 931 gcgaaggttt accgataag 19 <210> 932 <211> 20 <212> DNA <213> CNAG_02516 Southern blot probe primer <400> 932 tatggggtta ggagcgttgg 20 <210> 933 <211> 20 <212> DNA <213> NAT#210 STM primer <400> 933 ctagagcccg ccacaacgct 20 <210> 934 <211> 24 <212> DNA <213> STM common primer <400> 934 gcatgccctg cccctaagaa ttcg 24 <210> 935 <211> 19 <212> DNA <213> CNAG_04226 5 flanking region primer 1 <400> 935 gccctttttg gtatgacag 19 <210> 936 <211> 38 <212> DNA <213> CNAG_04226 5 flanking region primer 2 <400> 936 tcactggccg tcgttttact gtgagggatt ccttgttc 38 <210> 937 <211> 39 <212> DNA <213> CNAG_04226 3 flanking region primer 1 <400> 937 catggtcata gctgtttcct ggttggaaac aagcagtcg 39 <210> 938 <211> 20 <212> DNA <213> CNAG_04226 3 flanking region primer 2 <400> 938 tcagaagaat caagaggtcg 20 <210> 939 <211> 19 <212> DNA <213> CNAG_04226 diagnostic screening primer, pairing with B79 <400> 939 acttacattg ccttcctcg 19 <210> 940 <211> 19 <212> DNA <213> CNAG_04226 Southern blot probe primer <400> 940 atacatcagc ctgtcctgc 19 <210> 941 <211> 19 <212> DNA <213> CNAG_05170 5 flanking region primer 1 <400> 941 tgccttcgct ctgtaactc 19 <210> 942 <211> 34 <212> DNA <213> CNAG_05170 5 flanking region primer 2 <400> 942 ctggccgtcg ttttaccctt tttgaggctc gctg 34 <210> 943 <211> 36 <212> DNA <213> CNAG_05170 3 flanking region primer 1 <400> 943 gtcatagctg tttcctgaaa tgaaggagtg tgcggg 36 <210> 944 <211> 20 <212> DNA <213> CNAG_05170 3 flanking region primer 2 <400> 944 aataggacct caacaccagg 20 <210> 945 <211> 18 <212> DNA <213> CNAG_05170 diagnostic screening primer, pairing with B79 <400> 945 aacaagaagc ccctttcg 18 <210> 946 <211> 18 <212> DNA <213> CNAG_05170 Southern blot probe primer <400> 946 agtttttgga cccgagtg 18 <210> 947 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 947 ctttaaaggt ggtttgtg 18 <210> 948 <211> 24 <212> DNA <213> STM common primer <400> 948 gcatgccctg cccctaagaa ttcg 24 <210> 949 <211> 20 <212> DNA <213> CNAG_03902 5 flanking region primer 1 <400> 949 gcaaagtcaa gaaggtcagg 20 <210> 950 <211> 35 <212> DNA <213> CNAG_03902 5 flanking region primer 2 <400> 950 ctggccgtcg ttttacttct tctttggctc gtccg 35 <210> 951 <211> 39 <212> DNA <213> CNAG_03902 3 flanking region primer 1 <400> 951 gtcatagctg tttcctgggg agatgaaatg tgcgtttag 39 <210> 952 <211> 22 <212> DNA <213> CNAG_03902 3 flanking region primer 2 <400> 952 atgctttcgc cgtcatctcg tc 22 <210> 953 <211> 20 <212> DNA <213> CNAG_03902 diagnostic screening primer, pairing with B79 <400> 953 tgattacccc tgtcaaactc 20 <210> 954 <211> 19 <212> DNA <213> CNAG_03902 Southern blot probe primer <400> 954 cagaatcggg aagtaatgg 19 <210> 955 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 955 ctttaaaggt ggtttgtg 18 <210> 956 <211> 24 <212> DNA <213> STM common primer <400> 956 gcatgccctg cccctaagaa ttcg 24 <210> 957 <211> 20 <212> DNA <213> CNAG_06097 5 flanking region primer 1 <400> 957 cagtccgaag tttacaatcg 20 <210> 958 <211> 36 <212> DNA <213> CNAG_06097 5 flanking region primer 2 <400> 958 ctggccgtcg ttttactttc gtcgtgtgga gttggc 36 <210> 959 <211> 37 <212> DNA <213> CNAG_06097 3 flanking region primer 1 <400> 959 gtcatagctg tttcctgaga ttggagggtt tgagggc 37 <210> 960 <211> 19 <212> DNA <213> CNAG_06097 3 flanking region primer 2 <400> 960 tccaccgaag aaactgaag 19 <210> 961 <211> 20 <212> DNA <213> CNAG_06097 diagnostic screening primer, pairing with B79 <400> 961 aggtttgaga ggtttggaag 20 <210> 962 <211> 18 <212> DNA <213> CNAG_06097 Southern blot probe primer <400> 962 attatcggag cgttgtgg 18 <210> 963 <211> 20 <212> DNA <213> CNAG_03431 5 flanking region primer 1 <400> 963 aagcactctg tagcctgttg 20 <210> 964 <211> 39 <212> DNA <213> CNAG_03431 5 flanking region primer 2 <400> 964 gctcactggc cgtcgtttta ccttgaagat gatgcgtgc 39 <210> 965 <211> 40 <212> DNA <213> CNAG_03431 3 flanking region primer 1 <400> 965 catggtcata gctgtttcct gcgctcatag agaatgctgg 40 <210> 966 <211> 19 <212> DNA <213> CNAG_03431 3 flanking region primer 2 <400> 966 taggagtgtc tgccaaagc 19 <210> 967 <211> 19 <212> DNA <213> CNAG_03431 diagnostic screening primer, pairing with B79 <400> 967 tcatcctcat cccgaatac 19 <210> 968 <211> 19 <212> DNA <213> CNAG_03431 Southern blot probe primer <400> 968 actcttcgtg agacgcatc 19 <210> 969 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 969 accgacagct cgaacaagca agag 24 <210> 970 <211> 24 <212> DNA <213> STM common primer <400> 970 gcatgccctg cccctaagaa ttcg 24 <210> 971 <211> 21 <212> DNA <213> CNAG_00830 5 flanking region primer 1 <400> 971 ccgactaata ggtgtgctgt c 21 <210> 972 <211> 41 <212> DNA <213> CNAG_00830 5 flanking region primer 2 <400> 972 gctcactggc cgtcgtttta cgagatgaga tgagttgcgt g 41 <210> 973 <211> 41 <212> DNA <213> CNAG_00830 3 flanking region primer 1 <400> 973 catggtcata gctgtttcct gttgtgtccc catcactacc g 41 <210> 974 <211> 19 <212> DNA <213> CNAG_00830 3 flanking region primer 2 <400> 974 catccaaaga agcgaaacg 19 <210> 975 <211> 24 <212> DNA <213> CNAG_00830 diagnostic screening primer, pairing with B79 <400> 975 gaagagaagt agtggataga ctcg 24 <210> 976 <211> 19 <212> DNA <213> CNAG_00830 Southern blot probe primer <400> 976 tgatgccgta aatcgtagg 19 <210> 977 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 977 atttcgatgt atatatgtag c 21 <210> 978 <211> 24 <212> DNA <213> STM common primer <400> 978 gcatgccctg cccctaagaa ttcg 24 <210> 979 <211> 20 <212> DNA <213> CNAG_07443 5 flanking region primer 1 <400> 979 atctcttctc tcggcgtgtc 20 <210> 980 <211> 40 <212> DNA <213> CNAG_07443 5 flanking region primer 2 <400> 980 gctcactggc cgtcgtttta cacaggaaaa ggaggagtcg 40 <210> 981 <211> 40 <212> DNA <213> CNAG_07443 3 flanking region primer 1 <400> 981 catggtcata gctgtttcct gaagtctttc cgtctgaccc 40 <210> 982 <211> 18 <212> DNA <213> CNAG_07443 3 flanking region primer 2 <400> 982 tcgcatccct tgtattcg 18 <210> 983 <211> 19 <212> DNA <213> CNAG_07443 diagnostic screening primer, pairing with B79 <400> 983 tcaataatca gccgcctac 19 <210> 984 <211> 19 <212> DNA <213> CNAG_07443 Southern blot probe primer <400> 984 ttagattccg cataggtcc 19 <210> 985 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 985 acacctacat caaaccctcc c 21 <210> 986 <211> 24 <212> DNA <213> STM common primer <400> 986 gcatgccctg cccctaagaa ttcg 24 <210> 987 <211> 19 <212> DNA <213> CNAG_05186 5 flanking region primer 1 <400> 987 acaacggtca agaagggtc 19 <210> 988 <211> 39 <212> DNA <213> CNAG_05186 5 flanking region primer 2 <400> 988 gctcactggc cgtcgtttta caagtaaagg aacgcacgg 39 <210> 989 <211> 41 <212> DNA <213> CNAG_05186 3 flanking region primer 1 <400> 989 catggtcata gctgtttcct gcgcttgaga cgggagtaat c 41 <210> 990 <211> 20 <212> DNA <213> CNAG_05186 3 flanking region primer 2 <400> 990 gaggtaagga tgattggacc 20 <210> 991 <211> 23 <212> DNA <213> CNAG_05186 diagnostic screening primer, pairing with B79 <400> 991 gtgtgaagtg agtagtaacg agc 23 <210> 992 <211> 22 <212> DNA <213> CNAG_05186 Southern blot probe primer <400> 992 ttccctatct ctcttcatac cc 22 <210> 993 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 993 cgcccgccct cactatccac 20 <210> 994 <211> 24 <212> DNA <213> STM common primer <400> 994 gcatgccctg cccctaagaa ttcg 24 <210> 995 <211> 20 <212> DNA <213> CNAG_03561 5 flanking region primer 1 <400> 995 tgggcgagac gagatagaac 20 <210> 996 <211> 36 <212> DNA <213> CNAG_03561 5 flanking region primer 2 <400> 996 ctggccgtcg ttttacggat aaagatgatt cgggtc 36 <210> 997 <211> 36 <212> DNA <213> CNAG_03561 3 flanking region primer 1 <400> 997 gtcatagctg tttcctgtga tggctgattt cccgtg 36 <210> 998 <211> 36 <212> DNA <213> CNAG_03561 3 flanking region primer 2 <400> 998 gtcatagctg tttcctgtga tggctgattt cccgtg 36 <210> 999 <211> 20 <212> DNA <213> CNAG_03561 diagnostic screening primer, pairing with B79 <400> 999 aagaaagacg gtagcggtgg 20 <210> 1000 <211> 20 <212> DNA <213> CNAG_03561 Southern blot probe primer <400> 1000 ccacctcttc atccaatagc 20 <210> 1001 <211> 20 <212> DNA <213> NAT#43 STM primer <400> 1001 ccagctacca atcacgctac 20 <210> 1002 <211> 24 <212> DNA <213> STM common primer <400> 1002 gcatgccctg cccctaagaa ttcg 24 <210> 1003 <211> 19 <212> DNA <213> CNAG_04878 5 flanking region primer 1 <400> 1003 aagtccgtgc tggctaatc 19 <210> 1004 <211> 35 <212> DNA <213> CNAG_04878 5 flanking region primer 2 <400> 1004 ctggccgtcg ttttaccttt tcgctgcctt attcg 35 <210> 1005 <211> 41 <212> DNA <213> CNAG_04878 3 flanking region primer 1 <400> 1005 gtcatagctg tttcctgaaa acagggcata cggtgtcgtg g 41 <210> 1006 <211> 20 <212> DNA <213> CNAG_04878 3 flanking region primer 2 <400> 1006 gctgaccatc tcttccctac 20 <210> 1007 <211> 22 <212> DNA <213> CNAG_04878 diagnostic screening primer, pairing with B79 <400> 1007 ccttgtttgt gtctaccgta tc 22 <210> 1008 <211> 19 <212> DNA <213> CNAG_04878 Southern blot probe primer <400> 1008 agtgggaatc gtcgttgac 19 <210> 1009 <211> 20 <212> DNA <213> NAT#116 STM primer <400> 1009 gcacccaaga gctccatctc 20 <210> 1010 <211> 24 <212> DNA <213> STM common primer <400> 1010 gcatgccctg cccctaagaa ttcg 24 <210> 1011 <211> 19 <212> DNA <213> CNAG_03279 5 flanking region primer 1 <400> 1011 cgtttccagt tatcgcttc 19 <210> 1012 <211> 41 <212> DNA <213> CNAG_03279 5 flanking region primer 2 <400> 1012 gctcactggc cgtcgtttta cgcaatgacc cactttttga c 41 <210> 1013 <211> 41 <212> DNA <213> CNAG_03279 3 flanking region primer 1 <400> 1013 catggtcata gctgtttcct ggctttgcct gatgagtttt g 41 <210> 1014 <211> 19 <212> DNA <213> CNAG_03279 3 flanking region primer 2 <400> 1014 gtaacccact cggaagttg 19 <210> 1015 <211> 19 <212> DNA <213> CNAG_03279 diagnostic screening primer, pairing with B79 <400> 1015 ttgtctgtga ggtggaagg 19 <210> 1016 <211> 20 <212> DNA <213> CNAG_03279 Southern blot probe primer <400> 1016 caggctcgtt tgtagccttc 20 <210> 1017 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 1017 acagctccaa acctcgctaa acag 24 <210> 1018 <211> 24 <212> DNA <213> STM common primer <400> 1018 gcatgccctg cccctaagaa ttcg 24 <210> 1019 <211> 18 <212> DNA <213> CNAG_06921 5 flanking region primer 1 <400> 1019 gacaaagcaa caacagcg 18 <210> 1020 <211> 37 <212> DNA <213> CNAG_06921 5 flanking region primer 2 <400> 1020 tcactggccg tcgttttact cggatgacag atggttg 37 <210> 1021 <211> 39 <212> DNA <213> CNAG_06921 3 flanking region primer 1 <400> 1021 catggtcata gctgtttcct gaaagattgg gcgtctctg 39 <210> 1022 <211> 18 <212> DNA <213> CNAG_06921 3 flanking region primer 2 <400> 1022 atcatcagcg acatccag 18 <210> 1023 <211> 18 <212> DNA <213> CNAG_06921 diagnostic screening primer, pairing with B79 <400> 1023 aaagaacata cacgccgc 18 <210> 1024 <211> 18 <212> DNA <213> CNAG_06921 Southern blot probe primer <400> 1024 accatctgga tttctgcc 18 <210> 1025 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 1025 acgcaccaga cacacaacca g 21 <210> 1026 <211> 24 <212> DNA <213> STM common primer <400> 1026 gcatgccctg cccctaagaa ttcg 24 <210> 1027 <211> 20 <212> DNA <213> CNAG_02555 5 flanking region primer 1 <400> 1027 agaggaacac ggtatggtcg 20 <210> 1028 <211> 40 <212> DNA <213> CNAG_02555 5 flanking region primer 2 <400> 1028 gctcactggc cgtcgtttta ccaacaacgg tgaaccaaag 40 <210> 1029 <211> 41 <212> DNA <213> CNAG_02555 3 flanking region primer 1 <400> 1029 catggtcata gctgtttcct gcagagtgga ggtagaaacg g 41 <210> 1030 <211> 18 <212> DNA <213> CNAG_02555 3 flanking region primer 2 <400> 1030 tctgaaatgc tggctgtc 18 <210> 1031 <211> 20 <212> DNA <213> CNAG_02555 diagnostic screening primer, pairing with B79 <400> 1031 cccctcacaa cttgctaatg 20 <210> 1032 <211> 20 <212> DNA <213> CNAG_02555 Southern blot probe primer <400> 1032 ccgatgtcga cgtatgattg 20 <210> 1033 <211> 18 <212> DNA <213> NAT#270 STM primer <400> 1033 ggtggtttgt gtgggtag 18 <210> 1034 <211> 24 <212> DNA <213> STM common primer <400> 1034 gcatgccctg cccctaagaa ttcg 24 <210> 1035 <211> 19 <212> DNA <213> CNAG_03710 5 flanking region primer 1 <400> 1035 aagacaagaa gcagcagcg 19 <210> 1036 <211> 40 <212> DNA <213> CNAG_03710 5 flanking region primer 2 <400> 1036 gctcactggc cgtcgtttta ctatcattcc catccgtcgc 40 <210> 1037 <211> 42 <212> DNA <213> CNAG_03710 3 flanking region primer 1 <400> 1037 catggtcata gctgtttcct gcgctcaata acatcgtcag ac 42 <210> 1038 <211> 20 <212> DNA <213> CNAG_03710 3 flanking region primer 2 <400> 1038 ccagccttca tctctcttcc 20 <210> 1039 <211> 20 <212> DNA <213> CNAG_03710 diagnostic screening primer, pairing with B79 <400> 1039 cagaactgat aactggctgc 20 <210> 1040 <211> 20 <212> DNA <213> CNAG_03710 Southern blot probe primer <400> 1040 tcctctgcac caagactgtg 20 <210> 1041 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 1041 accgacagct cgaacaagca agag 24 <210> 1042 <211> 24 <212> DNA <213> STM common primer <400> 1042 gcatgccctg cccctaagaa ttcg 24 <210> 1043 <211> 20 <212> DNA <213> CNAG_03183 5 flanking region primer 1 <400> 1043 ctctatgagg gcaaagtcag 20 <210> 1044 <211> 40 <212> DNA <213> CNAG_03183 5 flanking region primer 2 <400> 1044 gctcactggc cgtcgtttta ctgttgttgc tgttggctcg 40 <210> 1045 <211> 40 <212> DNA <213> CNAG_03183 3 flanking region primer 1 <400> 1045 catggtcata gctgtttcct gttcttccaa tcccagccac 40 <210> 1046 <211> 19 <212> DNA <213> CNAG_03183 3 flanking region primer 2 <400> 1046 tacgcttcac agattacgc 19 <210> 1047 <211> 18 <212> DNA <213> CNAG_03183 diagnostic screening primer, pairing with B79 <400> 1047 tatacgtcca agacatcc 18 <210> 1048 <211> 20 <212> DNA <213> CNAG_03183 Southern blot probe primer <400> 1048 ctgattccga ggcttactcg 20 <210> 1049 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 1049 atttcgatgt atatatgtag c 21 <210> 1050 <211> 24 <212> DNA <213> STM common primer <400> 1050 gcatgccctg cccctaagaa ttcg 24 <210> 1051 <211> 20 <212> DNA <213> CNAG_04637 5 flanking region primer 1 <400> 1051 ccttcaagcc aaaatgagac 20 <210> 1052 <211> 40 <212> DNA <213> CNAG_04637 5 flanking region primer 2 <400> 1052 gctcactggc cgtcgtttta cgagatggag agaagcaacg 40 <210> 1053 <211> 43 <212> DNA <213> CNAG_04637 3 flanking region primer 1 <400> 1053 catggtcata gctgtttcct gggttgacgg atttactgaa gtc 43 <210> 1054 <211> 20 <212> DNA <213> CNAG_04637 3 flanking region primer 2 <400> 1054 tggaagatga ctcactcggg 20 <210> 1055 <211> 19 <212> DNA <213> CNAG_04637 diagnostic screening primer, pairing with B79 <400> 1055 tccatcaaag atacacccg 19 <210> 1056 <211> 20 <212> DNA <213> CNAG_04637 Southern blot probe primer <400> 1056 ttacatcttg aggcttagcg 20 <210> 1057 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 1057 cgcccgccct cactatccac 20 <210> 1058 <211> 24 <212> DNA <213> STM common primer <400> 1058 gcatgccctg cccctaagaa ttcg 24 <210> 1059 <211> 19 <212> DNA <213> CNAG_04398 5 flanking region primer 1 <400> 1059 agtctggaga aaaggggtc 19 <210> 1060 <211> 34 <212> DNA <213> CNAG_04398 5 flanking region primer 2 <400> 1060 ctggccgtcg ttttacgcaa acacacattc tcgg 34 <210> 1061 <211> 36 <212> DNA <213> CNAG_04398 3 flanking region primer 1 <400> 1061 gtcatagctg tttcctgttg gctaatgtag ggttgg 36 <210> 1062 <211> 18 <212> DNA <213> CNAG_04398 3 flanking region primer 2 <400> 1062 ccccttccca tttttcac 18 <210> 1063 <211> 22 <212> DNA <213> CNAG_04398 diagnostic screening primer, pairing with B79 <400> 1063 gccactttct gatttccgta tc 22 <210> 1064 <211> 18 <212> DNA <213> CNAG_04398 Southern blot probe primer <400> 1064 tatttgctgc ctgaccac 18 <210> 1065 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 1065 ccatagaact agctaaagca 20 <210> 1066 <211> 24 <212> DNA <213> STM common primer <400> 1066 gcatgccctg cccctaagaa ttcg 24 <210> 1067 <211> 20 <212> DNA <213> CNAG_01454 5 flanking region primer 1 <400> 1067 ggaagtttgg aggaggatag 20 <210> 1068 <211> 37 <212> DNA <213> CNAG_01454 5 flanking region primer 2 <400> 1068 tcactggccg tcgttttacc agattcgtgg tcaaagc 37 <210> 1069 <211> 40 <212> DNA <213> CNAG_01454 3 flanking region primer 1 <400> 1069 catggtcata gctgtttcct ggccttgaat gatgaccagc 40 <210> 1070 <211> 18 <212> DNA <213> CNAG_01454 3 flanking region primer 2 <400> 1070 cggatttgag atgatgcc 18 <210> 1071 <211> 22 <212> DNA <213> CNAG_01454 diagnostic screening primer, pairing with B79 <400> 1071 gttagaggag tcattctttg gc 22 <210> 1072 <211> 20 <212> DNA <213> CNAG_01454 Southern blot probe primer <400> 1072 caaaaggtca aggaaaggtg 20 <210> 1073 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 1073 cgcaaaatca ctagccctat agcg 24 <210> 1074 <211> 24 <212> DNA <213> STM common primer <400> 1074 gcatgccctg cccctaagaa ttcg 24 <210> 1075 <211> 18 <212> DNA <213> CNAG_04268 5 flanking region primer 1 <400> 1075 tcgtctttgt gtggttgc 18 <210> 1076 <211> 38 <212> DNA <213> CNAG_04268 5 flanking region primer 2 <400> 1076 tcactggccg tcgttttaca cttatggcga cttacggg 38 <210> 1077 <211> 40 <212> DNA <213> CNAG_04268 3 flanking region primer 1 <400> 1077 catggtcata gctgtttcct ggcaatgtct gtatcaccgc 40 <210> 1078 <211> 19 <212> DNA <213> CNAG_04268 3 flanking region primer 2 <400> 1078 cgtccgcctg tattttatc 19 <210> 1079 <211> 18 <212> DNA <213> CNAG_04268 diagnostic screening primer, pairing with B79 <400> 1079 atgatgctgt tttgcccc 18 <210> 1080 <211> 18 <212> DNA <213> CNAG_04268 Southern blot probe primer <400> 1080 ttctttccac atcggtcc 18 <210> 1081 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 1081 ccatagcgat atctacccca atct 24 <210> 1082 <211> 24 <212> DNA <213> STM common primer <400> 1082 gcatgccctg cccctaagaa ttcg 24 <210> 1083 <211> 22 <212> DNA <213> CNAG_00031 5 flanking region primer 1 <400> 1083 atccaatcat cggcttccca gc 22 <210> 1084 <211> 35 <212> DNA <213> CNAG_00031 5 flanking region primer 2 <400> 1084 ctggccgtcg ttttacactt gaactcccat ctgcg 35 <210> 1085 <211> 37 <212> DNA <213> CNAG_00031 3 flanking region primer 1 <400> 1085 gtcatagctg tttcctggaa atgcgatgaa tgggttg 37 <210> 1086 <211> 21 <212> DNA <213> CNAG_00031 3 flanking region primer 2 <400> 1086 cctcttgccc taatgagata g 21 <210> 1087 <211> 18 <212> DNA <213> CNAG_00031 diagnostic screening primer, pairing with B79 <400> 1087 ccgtgttcgc aatctatg 18 <210> 1088 <211> 19 <212> DNA <213> CNAG_00031 Southern blot probe primer <400> 1088 gaatcagcat ttgggttcg 19 <210> 1089 <211> 20 <212> DNA <213> NAT#116 STM primer <400> 1089 gcacccaaga gctccatctc 20 <210> 1090 <211> 24 <212> DNA <213> STM common primer <400> 1090 gcatgccctg cccctaagaa ttcg 24 <210> 1091 <211> 19 <212> DNA <213> CNAG_07922 5 flanking region primer 1 <400> 1091 cctcttccgc tattttgtg 19 <210> 1092 <211> 39 <212> DNA <213> CNAG_07922 5 flanking region primer 2 <400> 1092 gctcactggc cgtcgtttta ccattgctgt tgtgggaac 39 <210> 1093 <211> 41 <212> DNA <213> CNAG_07922 3 flanking region primer 1 <400> 1093 catggtcata gctgtttcct gaagtggagt tggaagaggc g 41 <210> 1094 <211> 22 <212> DNA <213> CNAG_07922 3 flanking region primer 2 <400> 1094 caactatcca atacctgttc cg 22 <210> 1095 <211> 19 <212> DNA <213> CNAG_07922 diagnostic screening primer, pairing with B79 <400> 1095 cacggcttgt cttgaagtc 19 <210> 1096 <211> 18 <212> DNA <213> CNAG_07922 Southern blot probe primer <400> 1096 ttgacggttt tggcacag 18 <210> 1097 <211> 20 <212> DNA <213> NAT#116 STM primer <400> 1097 gcacccaaga gctccatctc 20 <210> 1098 <211> 24 <212> DNA <213> STM common primer <400> 1098 gcatgccctg cccctaagaa ttcg 24 <210> 1099 <211> 19 <212> DNA <213> CNAG_04841 5 flanking region primer 1 <400> 1099 tgaggtgttc cgtgtattg 19 <210> 1100 <211> 35 <212> DNA <213> CNAG_04841 5 flanking region primer 2 <400> 1100 ctggccgtcg ttttacctct tgactggtaa tgggg 35 <210> 1101 <211> 35 <212> DNA <213> CNAG_04841 3 flanking region primer 1 <400> 1101 gtcatagctg tttcctgatt cccaaggtga cgaag 35 <210> 1102 <211> 20 <212> DNA <213> CNAG_04841 3 flanking region primer 2 <400> 1102 tctccaataa gttcaggcac 20 <210> 1103 <211> 22 <212> DNA <213> CNAG_04841 diagnostic screening primer, pairing with B79 <400> 1103 taaagacgga ggtgggagta gg 22 <210> 1104 <211> 18 <212> DNA <213> CNAG_04841 Southern blot probe primer <400> 1104 tgagattggg aaagagcg 18 <210> 1105 <211> 24 <212> DNA <213> NAT#119 STM primer <400> 1105 ctccccacat aaagagagct aaac 24 <210> 1106 <211> 24 <212> DNA <213> STM common primer <400> 1106 gcatgccctg cccctaagaa ttcg 24 <210> 1107 <211> 20 <212> DNA <213> CNAG_01948 5 flanking region primer 1 <400> 1107 cttggtaaat ccgtattccg 20 <210> 1108 <211> 35 <212> DNA <213> CNAG_01948 5 flanking region primer 2 <400> 1108 ctggccgtcg ttttaccctt tcgcttcaac ctacg 35 <210> 1109 <211> 35 <212> DNA <213> CNAG_01948 3 flanking region primer 1 <400> 1109 gtcatagctg tttcctgtga tttggcgact ctgag 35 <210> 1110 <211> 20 <212> DNA <213> CNAG_01948 3 flanking region primer 2 <400> 1110 tgtgtttctc atcgtaagcg 20 <210> 1111 <211> 22 <212> DNA <213> CNAG_01948 diagnostic screening primer, pairing with B79 <400> 1111 ttgacttccc aggttacact ac 22 <210> 1112 <211> 19 <212> DNA <213> CNAG_01948 Southern blot probe primer <400> 1112 cattgggact cttgtaggg 19 <210> 1113 <211> 24 <212> DNA <213> NAT#119 STM primer <400> 1113 ctccccacat aaagagagct aaac 24 <210> 1114 <211> 24 <212> DNA <213> STM common primer <400> 1114 gcatgccctg cccctaagaa ttcg 24 <210> 1115 <211> 19 <212> DNA <213> CNAG_07435 5 flanking region primer 1 <400> 1115 caaccttcaa agtcttgcc 19 <210> 1116 <211> 39 <212> DNA <213> CNAG_07435 5 flanking region primer 2 <400> 1116 tcactggccg tcgttttaca acgaggagtc atactgtgc 39 <210> 1117 <211> 41 <212> DNA <213> CNAG_07435 3 flanking region primer 1 <400> 1117 catggtcata gctgtttcct gaaaggcgtc aaagggcgaa g 41 <210> 1118 <211> 20 <212> DNA <213> CNAG_07435 3 flanking region primer 2 <400> 1118 ggtttggagc gtattgattg 20 <210> 1119 <211> 19 <212> DNA <213> CNAG_07435 diagnostic screening primer, pairing with B79 <400> 1119 aagagcggca ttgaatcgg 19 <210> 1120 <211> 20 <212> DNA <213> CNAG_07435 Southern blot probe primer <400> 1120 gcgtcacttt ggacatactc 20 <210> 1121 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 1121 ctatcgacca accaacacag 20 <210> 1122 <211> 24 <212> DNA <213> STM common primer <400> 1122 gcatgccctg cccctaagaa ttcg 24 <210> 1123 <211> 22 <212> DNA <213> CNAG_00505 5 flanking region primer 1 <400> 1123 tttcaggcac caacaggcac ag 22 <210> 1124 <211> 38 <212> DNA <213> CNAG_00505 5 flanking region primer 2 <400> 1124 ctggccgtcg ttttacttcc gatggtgccc ttgagtag 38 <210> 1125 <211> 38 <212> DNA <213> CNAG_00505 3 flanking region primer 1 <400> 1125 gtcatagctg tttcctgggt cttccaaatg tcacagtc 38 <210> 1126 <211> 19 <212> DNA <213> CNAG_00505 3 flanking region primer 2 <400> 1126 aaagaagaga cgcctcacg 19 <210> 1127 <211> 20 <212> DNA <213> CNAG_00505 diagnostic screening primer, pairing with B79 <400> 1127 tgatagacac cacggcagtc 20 <210> 1128 <211> 20 <212> DNA <213> CNAG_00505 Southern blot probe primer <400> 1128 cgaggattgt caaatgaagg 20 <210> 1129 <211> 24 <212> DNA <213> NAT#125 STM primer <400> 1129 cgctacagcc agcgcgcgca agcg 24 <210> 1130 <211> 24 <212> DNA <213> STM common primer <400> 1130 gcatgccctg cccctaagaa ttcg 24 <210> 1131 <211> 18 <212> DNA <213> CNAG_03115 5 flanking region primer 1 <400> 1131 cgaaggaagt aaaacggg 18 <210> 1132 <211> 37 <212> DNA <213> CNAG_03115 5 flanking region primer 2 <400> 1132 ctggccgtcg ttttacaagc actcaccact ctcctac 37 <210> 1133 <211> 36 <212> DNA <213> CNAG_03115 3 flanking region primer 1 <400> 1133 gtcatagctg tttcctgtgc ttggatagac gacgag 36 <210> 1134 <211> 20 <212> DNA <213> CNAG_03115 3 flanking region primer 2 <400> 1134 tcaatcaaag cgttcggggg 20 <210> 1135 <211> 20 <212> DNA <213> CNAG_03115 diagnostic screening primer, pairing with B79 <400> 1135 aatccgactt ctcagccctg 20 <210> 1136 <211> 20 <212> DNA <213> CNAG_03115 Southern blot probe primer <400> 1136 atctgaggga agggctacag 20 <210> 1137 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 1137 caccaactcc ccatctccat 20 <210> 1138 <211> 24 <212> DNA <213> STM common primer <400> 1138 gcatgccctg cccctaagaa ttcg 24 <210> 1139 <211> 19 <212> DNA <213> CNAG_04807 5 flanking region primer 1 <400> 1139 gcgtttcctc gtgaacttc 19 <210> 1140 <211> 37 <212> DNA <213> CNAG_04807 5 flanking region primer 2 <400> 1140 ctggccgtcg ttttacggaa gaagggttga gacactc 37 <210> 1141 <211> 36 <212> DNA <213> CNAG_04807 3 flanking region primer 1 <400> 1141 gtcatagctg tttcctgggc aagcataatg gcaaag 36 <210> 1142 <211> 18 <212> DNA <213> CNAG_04807 3 flanking region primer 2 <400> 1142 ggatttccga agccaaac 18 <210> 1143 <211> 19 <212> DNA <213> CNAG_04807 diagnostic screening primer, pairing with B79 <400> 1143 tggcacagtt caatgttcc 19 <210> 1144 <211> 18 <212> DNA <213> CNAG_04807 Southern blot probe primer <400> 1144 cgacaataca aacacgcc 18 <210> 1145 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 1145 caccaactcc ccatctccat 20 <210> 1146 <211> 24 <212> DNA <213> STM common primer <400> 1146 gcatgccctg cccctaagaa ttcg 24 <210> 1147 <211> 19 <212> DNA <213> CNAG_02435 5 flanking region primer 1 <400> 1147 atcgttagca gggtgagtc 19 <210> 1148 <211> 42 <212> DNA <213> CNAG_02435 5 flanking region primer 2 <400> 1148 gctcactggc cgtcgtttta ctcctattca gggactatca cc 42 <210> 1149 <211> 39 <212> DNA <213> CNAG_02435 3 flanking region primer 1 <400> 1149 catggtcata gctgtttcct gacctatttt ccacgcagc 39 <210> 1150 <211> 18 <212> DNA <213> CNAG_02435 3 flanking region primer 2 <400> 1150 tcagcctcgc acataatc 18 <210> 1151 <211> 18 <212> DNA <213> CNAG_02435 diagnostic screening primer, pairing with B79 <400> 1151 cgagccgaaa agaacaag 18 <210> 1152 <211> 20 <212> DNA <213> CNAG_02435 Southern blot probe primer <400> 1152 gattatctgt ttggttcggc 20 <210> 1153 <211> 24 <212> DNA <213> NAT#184 STM primer <400> 1153 atatatggct cgagctagat agag 24 <210> 1154 <211> 24 <212> DNA <213> STM common primer <400> 1154 gcatgccctg cccctaagaa ttcg 24 <210> 1155 <211> 20 <212> DNA <213> CNAG_02364 5 flanking region primer 1 <400> 1155 gaagtcaaca ggaagaagcc 20 <210> 1156 <211> 34 <212> DNA <213> CNAG_02364 5 flanking region primer 2 <400> 1156 ctggccgtcg ttttactttg cgtgaaggtg tagg 34 <210> 1157 <211> 36 <212> DNA <213> CNAG_02364 3 flanking region primer 1 <400> 1157 gtcatagctg tttcctgtgc tcatcttcac ggctac 36 <210> 1158 <211> 19 <212> DNA <213> CNAG_02364 3 flanking region primer 2 <400> 1158 cattatccgc ctgatgttg 19 <210> 1159 <211> 19 <212> DNA <213> CNAG_02364 diagnostic screening primer, pairing with B79 <400> 1159 gcacttccaa caaagccac 19 <210> 1160 <211> 19 <212> DNA <213> CNAG_02364 Southern blot probe primer <400> 1160 gactttttgc gtgaaggtg 19 <210> 1161 <211> 24 <212> DNA <213> NAT#184 STM primer <400> 1161 atatatggct cgagctagat agag 24 <210> 1162 <211> 24 <212> DNA <213> STM common primer <400> 1162 gcatgccctg cccctaagaa ttcg 24 <210> 1163 <211> 20 <212> DNA <213> CNAG_05420 5 flanking region primer 1 <400> 1163 cgcctttgag gtattcactc 20 <210> 1164 <211> 39 <212> DNA <213> CNAG_05420 5 flanking region primer 2 <400> 1164 tcactggccg tcgttttacg gtgttggtca cgaatctac 39 <210> 1165 <211> 40 <212> DNA <213> CNAG_05420 3 flanking region primer 1 <400> 1165 catggtcata gctgtttcct gcggctttac gaaatgagtg 40 <210> 1166 <211> 21 <212> DNA <213> CNAG_05420 3 flanking region primer 2 <400> 1166 cctgaaagac atagcgacaa g 21 <210> 1167 <211> 19 <212> DNA <213> CNAG_05420 diagnostic screening primer, pairing with B79 <400> 1167 atcctgtctt gctgtggtc 19 <210> 1168 <211> 18 <212> DNA <213> CNAG_05420 Southern blot probe primer <400> 1168 tccagcatcg tcattgag 18 <210> 1169 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 1169 atatggatgt ttttagcgag 20 <210> 1170 <211> 24 <212> DNA <213> STM common primer <400> 1170 gcatgccctg cccctaagaa ttcg 24 <210> 1171 <211> 19 <212> DNA <213> CNAG_05093 5 flanking region primer 1 <400> 1171 aatgtattgc caccgtgcc 19 <210> 1172 <211> 41 <212> DNA <213> CNAG_05093 5 flanking region primer 2 <400> 1172 tcactggccg tcgttttact tggggtgatg tgttctcgtc c 41 <210> 1173 <211> 41 <212> DNA <213> CNAG_05093 3 flanking region primer 1 <400> 1173 catggtcata gctgtttcct gacaggcaga ttgggaagtg c 41 <210> 1174 <211> 19 <212> DNA <213> CNAG_05093 3 flanking region primer 2 <400> 1174 tgtatgccgt gggaacaag 19 <210> 1175 <211> 24 <212> DNA <213> CNAG_05093 diagnostic screening primer, pairing with B79 <400> 1175 cgaagtctca aacctatcta ctcg 24 <210> 1176 <211> 22 <212> DNA <213> CNAG_05093 Southern blot probe primer <400> 1176 ttgctgaggc gaacttgttg gc 22 <210> 1177 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 1177 caccctctat ctcgagaaag ctcc 24 <210> 1178 <211> 24 <212> DNA <213> STM common primer <400> 1178 gcatgccctg cccctaagaa ttcg 24 <210> 1179 <211> 19 <212> DNA <213> CNAG_06814 5 flanking region primer 1 <400> 1179 tgggaatggc aaaaggcac 19 <210> 1180 <211> 35 <212> DNA <213> CNAG_06814 5 flanking region primer 2 <400> 1180 ctggccgtcg ttttacggta ttttcgtcca gtaaa 35 <210> 1181 <211> 35 <212> DNA <213> CNAG_06814 3 flanking region primer 1 <400> 1181 gtcatagctg tttcctgtgc agcctatgcc tcctt 35 <210> 1182 <211> 19 <212> DNA <213> CNAG_06814 3 flanking region primer 2 <400> 1182 taagtgtgtt cacggtgcg 19 <210> 1183 <211> 19 <212> DNA <213> CNAG_06814 diagnostic screening primer, pairing with B79 <400> 1183 tggttaccga agtagggag 19 <210> 1184 <211> 19 <212> DNA <213> CNAG_06814 Southern blot probe primer <400> 1184 atttggaggt ggagttctg 19 <210> 1185 <211> 22 <212> DNA <213> NAT#208 STM primer <400> 1185 tggtcgcggg agatcgtggt tt 22 <210> 1186 <211> 24 <212> DNA <213> STM common primer <400> 1186 gcatgccctg cccctaagaa ttcg 24 <210> 1187 <211> 21 <212> DNA <213> CNAG_05431 5 flanking region primer 1 <400> 1187 catcagtctt gcttcttctg c 21 <210> 1188 <211> 40 <212> DNA <213> CNAG_05431 5 flanking region primer 2 <400> 1188 tcactggccg tcgttttaca gagtcagaat aggtgttggg 40 <210> 1189 <211> 40 <212> DNA <213> CNAG_05431 3 flanking region primer 1 <400> 1189 catggtcata gctgtttcct gagggagagg attgaagtgg 40 <210> 1190 <211> 20 <212> DNA <213> CNAG_05431 3 flanking region primer 2 <400> 1190 cattactttg actgggaggg 20 <210> 1191 <211> 19 <212> DNA <213> CNAG_05431 diagnostic screening primer, pairing with B79 <400> 1191 cacggcaact tatgctctc 19 <210> 1192 <211> 20 <212> DNA <213> CNAG_05431 Southern blot probe primer <400> 1192 actcggtgtt ggtgaaacgg 20 <210> 1193 <211> 22 <212> DNA <213> NAT#208 STM primer <400> 1193 tggtcgcggg agatcgtggt tt 22 <210> 1194 <211> 24 <212> DNA <213> STM common primer <400> 1194 gcatgccctg cccctaagaa ttcg 24 <210> 1195 <211> 20 <212> DNA <213> CNAG_05642 5 flanking region primer 1 <400> 1195 agagggataa gagcctgacg 20 <210> 1196 <211> 38 <212> DNA <213> CNAG_05642 5 flanking region primer 2 <400> 1196 tcactggccg tcgttttaca gaaggaagtt gggaagcg 38 <210> 1197 <211> 41 <212> DNA <213> CNAG_05642 3 flanking region primer 1 <400> 1197 catggtcata gctgtttcct gatgattaca gaggggagga c 41 <210> 1198 <211> 19 <212> DNA <213> CNAG_05642 3 flanking region primer 2 <400> 1198 tgtatcccaa gcagcaaag 19 <210> 1199 <211> 20 <212> DNA <213> CNAG_05642 diagnostic screening primer, pairing with B79 <400> 1199 ctcggaaata aatgtggagc 20 <210> 1200 <211> 20 <212> DNA <213> CNAG_05642 Southern blot probe primer <400> 1200 tacctgccag tgcttcaacc 20 <210> 1201 <211> 20 <212> DNA <213> NAT#210 STM primer <400> 1201 ctagagcccg ccacaacgct 20 <210> 1202 <211> 24 <212> DNA <213> STM common primer <400> 1202 gcatgccctg cccctaagaa ttcg 24 <210> 1203 <211> 19 <212> DNA <213> CNAG_03212 5 flanking region primer 1 <400> 1203 gacacccaaa caagcaaac 19 <210> 1204 <211> 39 <212> DNA <213> CNAG_03212 5 flanking region primer 2 <400> 1204 gctcactggc cgtcgtttta cctggggtgt ttgttttcg 39 <210> 1205 <211> 41 <212> DNA <213> CNAG_03212 3 flanking region primer 1 <400> 1205 catggtcata gctgtttcct ggggttcaga ttttggatac c 41 <210> 1206 <211> 19 <212> DNA <213> CNAG_03212 3 flanking region primer 2 <400> 1206 ggcgaggtgt tgtaaatcc 19 <210> 1207 <211> 22 <212> DNA <213> CNAG_03212 diagnostic screening primer, pairing with B79 <400> 1207 gcgtatctgg catcttgtta tc 22 <210> 1208 <211> 18 <212> DNA <213> CNAG_03212 Southern blot probe primer <400> 1208 tgaggaatgc caatggtg 18 <210> 1209 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 1209 gcggtcgctt tatagcgatt 20 <210> 1210 <211> 24 <212> DNA <213> STM common primer <400> 1210 gcatgccctg cccctaagaa ttcg 24 <210> 1211 <211> 18 <212> DNA <213> CNAG_03212 5 flanking region primer 1 <400> 1211 gcgaaacttt gtgagtgc 18 <210> 1212 <211> 40 <212> DNA <213> CNAG_03212 5 flanking region primer 2 <400> 1212 gctcactggc cgtcgtttta ccaagcagca agatggagtc 40 <210> 1213 <211> 40 <212> DNA <213> CNAG_03212 3 flanking region primer 1 <400> 1213 catggtcata gctgtttcct gtcgtaagtt cggtgtcagc 40 <210> 1214 <211> 20 <212> DNA <213> CNAG_03212 3 flanking region primer 2 <400> 1214 aagtcttatt atgggcgagg 20 <210> 1215 <211> 19 <212> DNA <213> CNAG_03212 diagnostic screening primer, pairing with B79 <400> 1215 tgtcccgttg ataggtgtc 19 <210> 1216 <211> 20 <212> DNA <213> CNAG_03212 Southern blot probe primer <400> 1216 tcctctccct cctattcagg 20 <210> 1217 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 1217 gcggtcgctt tatagcgatt 20 <210> 1218 <211> 24 <212> DNA <213> STM common primer <400> 1218 gcatgccctg cccctaagaa ttcg 24 <210> 1219 <211> 19 <212> DNA <213> CNAG_07924 5 flanking region primer 1 <400> 1219 acctaatcag gaaagcccc 19 <210> 1220 <211> 39 <212> DNA <213> CNAG_07924 5 flanking region primer 2 <400> 1220 tcactggccg tcgttttacc attccttctc cgaaatctc 39 <210> 1221 <211> 40 <212> DNA <213> CNAG_07924 3 flanking region primer 1 <400> 1221 catggtcata gctgtttcct gttatcctcc cgaataccgc 40 <210> 1222 <211> 18 <212> DNA <213> CNAG_07924 3 flanking region primer 2 <400> 1222 ccgcaccaaa ctatttcg 18 <210> 1223 <211> 18 <212> DNA <213> CNAG_07924 diagnostic screening primer, pairing with B79 <400> 1223 atcgcacgaa aggcaaag 18 <210> 1224 <211> 20 <212> DNA <213> CNAG_07924 Southern blot probe primer <400> 1224 tctaaacccg tctctgactg 20 <210> 1225 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 1225 ctccacatcc atcgctccaa 20 <210> 1226 <211> 24 <212> DNA <213> STM common primer <400> 1226 gcatgccctg cccctaagaa ttcg 24 <210> 1227 <211> 22 <212> DNA <213> CNAG_00156 5 flanking region primer 1 <400> 1227 cccgattttg ctttttagtg cg 22 <210> 1228 <211> 36 <212> DNA <213> CNAG_00156 5 flanking region primer 2 <400> 1228 ctggccgtcg ttttacttgc cgtctttgct tgggtc 36 <210> 1229 <211> 36 <212> DNA <213> CNAG_00156 3 flanking region primer 1 <400> 1229 gtcatagctg tttcctgatc aagtccactc ccgaag 36 <210> 1230 <211> 19 <212> DNA <213> CNAG_00156 3 flanking region primer <400> 1230 tagatgatga gggcgaacg 19 <210> 1231 <211> 20 <212> DNA <213> CNAG_00156 diagnostic screening primer, pairing with B79 <400> 1231 gcttcgttgg ttcgttagtc 20 <210> 1232 <211> 20 <212> DNA <213> CNAG_00156 Southern blot probe primer <400> 1232 caatccgttt ctctcacctc 20 <210> 1233 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 1233 gagagatccc aacatcacgc 20 <210> 1234 <211> 24 <212> DNA <213> STM common primer <400> 1234 gcatgccctg cccctaagaa ttcg 24 <210> 1235 <211> 19 <212> DNA <213> CNAG_01173 5 flanking region primer 1 <400> 1235 agtatcccaa gcatccctc 19 <210> 1236 <211> 39 <212> DNA <213> CNAG_01173 5 flanking region primer 2 <400> 1236 tcactggccg tcgttttacg gcaggagcaa agtgtattc 39 <210> 1237 <211> 39 <212> DNA <213> CNAG_01173 3 flanking region primer 1 <400> 1237 catggtcata gctgtttcct gtatggtggc atctgaacc 39 <210> 1238 <211> 20 <212> DNA <213> CNAG_01173 3 flanking region primer 2 <400> 1238 ctgacaagtt taccaggacg 20 <210> 1239 <211> 20 <212> DNA <213> CNAG_01173 diagnostic screening primer, pairing with B79 <400> 1239 gcttctgtgt gtctcctctg 20 <210> 1240 <211> 20 <212> DNA <213> CNAG_01173 Southern blot probe primer <400> 1240 gagaaagaag gattccgaag 20 <210> 1241 <211> 24 <212> DNA <213> NAT#242 STM primer <400> 1241 gtagcgatag gggtgtcgct ttag 24 <210> 1242 <211> 24 <212> DNA <213> STM common primer <400> 1242 gcatgccctg cccctaagaa ttcg 24 <210> 1243 <211> 19 <212> DNA <213> CNAG_06871 5 flanking region primer 1 <400> 1243 tcaggatggg agtgtttgc 19 <210> 1244 <211> 37 <212> DNA <213> CNAG_06871 5 flanking region primer 2 <400> 1244 tcactggccg tcgttttacc cttctccttt tgcttgg 37 <210> 1245 <211> 41 <212> DNA <213> CNAG_06871 3 flanking region primer 1 <400> 1245 catggtcata gctgtttcct gcgctcttat taccgcttca g 41 <210> 1246 <211> 20 <212> DNA <213> CNAG_06871 3 flanking region primer 2 <400> 1246 tcggtatttc agtagggatg 20 <210> 1247 <211> 19 <212> DNA <213> CNAG_06871 diagnostic screening primer, pairing with B79 <400> 1247 cgactgctgg atgtatgag 19 <210> 1248 <211> 18 <212> DNA <213> CNAG_06871 Southern blot probe primer <400> 1248 cttgactgcc tcatctgc 18 <210> 1249 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 1249 acacctacat caaaccctcc c 21 <210> 1250 <211> 24 <212> DNA <213> STM common primer <400> 1250 gcatgccctg cccctaagaa ttcg 24 <210> 1251 <211> 18 <212> DNA <213> CNAG_04774 5 flanking region primer 1 <400> 1251 atttgaatag ccgcaccc 18 <210> 1252 <211> 35 <212> DNA <213> CNAG_04774 5 flanking region primer 2 <400> 1252 ctggccgtcg ttttacattg ggggatgctg gaaag 35 <210> 1253 <211> 36 <212> DNA <213> CNAG_04774 3 flanking region primer 1 <400> 1253 gtcatagctg tttcctgggt ggactgccaa actatg 36 <210> 1254 <211> 19 <212> DNA <213> CNAG_04774 3 flanking region primer 2 <400> 1254 aggcatttct gttcctccg 19 <210> 1255 <211> 19 <212> DNA <213> CNAG_04774 diagnostic screening primer, pairing with B79 <400> 1255 cgatagacgg aggatgaag 19 <210> 1256 <211> 18 <212> DNA <213> CNAG_04774 Southern blot probe primer <400> 1256 atcgttgctt ccgcttag 18 <210> 1257 <211> 21 <212> DNA <213> NAT#146 STM primer <400> 1257 actagccccc cctcaccacc t 21 <210> 1258 <211> 24 <212> DNA <213> STM common primer <400> 1258 gcatgccctg cccctaagaa ttcg 24 <210> 1259 <211> 18 <212> DNA <213> CNAG_00841 5 flanking region primer 1 <400> 1259 tcggcaagtt tagcaagc 18 <210> 1260 <211> 39 <212> DNA <213> CNAG_00841 5 flanking region primer 2 <400> 1260 gctcactggc cgtcgtttta cttatttctg ggctgtcgc 39 <210> 1261 <211> 40 <212> DNA <213> CNAG_00841 3 flanking region primer 1 <400> 1261 catggtcata gctgtttcct gtctgtggag tcagccaaac 40 <210> 1262 <211> 23 <212> DNA <213> CNAG_00841 3 flanking region primer 2 <400> 1262 cggtatcctc gtgagtctat ttc 23 <210> 1263 <211> 21 <212> DNA <213> CNAG_00841 diagnostic screening primer, pairing with B79 <400> 1263 gcagttgctg aagtgaaaga c 21 <210> 1264 <211> 20 <212> DNA <213> CNAG_00841 Southern blot probe primer <400> 1264 tgggatagaa gaaggctgtg 20 <210> 1265 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 1265 tatccccctc tccgctctct agca 24 <210> 1266 <211> 24 <212> DNA <213> STM common primer <400> 1266 gcatgccctg cccctaagaa ttcg 24 <210> 1267 <211> 18 <212> DNA <213> CNAG_04836 5 flanking region primer 1 <400> 1267 tagaaaagcc acaacccc 18 <210> 1268 <211> 37 <212> DNA <213> CNAG_04836 5 flanking region primer 2 <400> 1268 tcactggccg tcgttttacg agtttctttt ggggtgg 37 <210> 1269 <211> 40 <212> DNA <213> CNAG_04836 3 flanking region primer 1 <400> 1269 catggtcata gctgtttcct gacgagaagg ggaatgatgg 40 <210> 1270 <211> 19 <212> DNA <213> CNAG_04836 3 flanking region primer 2 <400> 1270 tgggcatctt gctcattac 19 <210> 1271 <211> 19 <212> DNA <213> CNAG_04836 diagnostic screening primer, pairing with B79 <400> 1271 cggagagcaa atagcacag 19 <210> 1272 <211> 20 <212> DNA <213> CNAG_04836 Southern blot probe primer <400> 1272 aggtgtcgta tcgtatttgc 20 <210> 1273 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 1273 ctatcgacca accaacacag 20 <210> 1274 <211> 24 <212> DNA <213> STM common primer <400> 1274 gcatgccctg cccctaagaa ttcg 24 <210> 1275 <211> 19 <212> DNA <213> CNAG_02603 5 flanking region primer 1 <400> 1275 tcaaaacgcc tttcggaac 19 <210> 1276 <211> 40 <212> DNA <213> CNAG_02603 5 flanking region primer 2 <400> 1276 gctcactggc cgtcgtttta cacaaatggc agttccctcg 40 <210> 1277 <211> 43 <212> DNA <213> CNAG_02603 3 flanking region primer 1 <400> 1277 catggtcata gctgtttcct gatctctccc tcacttcagt cgg 43 <210> 1278 <211> 19 <212> DNA <213> CNAG_02603 3 flanking region primer 2 <400> 1278 taggaccaac ttcagaccg 19 <210> 1279 <211> 20 <212> DNA <213> CNAG_02603 diagnostic screening primer, pairing with B79 <400> 1279 gcctatggaa gaagacaacg 20 <210> 1280 <211> 20 <212> DNA <213> CNAG_02603 Southern blot probe primer <400> 1280 gtcgtgtagt tttgaatggc 20 <210> 1281 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 1281 aacctttaaa tgggtagag 19 <210> 1282 <211> 24 <212> DNA <213> STM common primer <400> 1282 gcatgccctg cccctaagaa ttcg 24 <210> 1283 <211> 20 <212> DNA <213> CNAG_03059 5 flanking region primer 1 <400> 1283 aatggcgtta ccgacgagag 20 <210> 1284 <211> 36 <212> DNA <213> CNAG_03059 5 flanking region primer 2 <400> 1284 ctggccgtcg ttttacggta ttgagagtgc cgagtg 36 <210> 1285 <211> 38 <212> DNA <213> CNAG_03059 3 flanking region primer 1 <400> 1285 gtcatagctg tttcctgcag actccctctt cttacctg 38 <210> 1286 <211> 24 <212> DNA <213> CNAG_03059 3 flanking region primer 2 <400> 1286 cgttactgcg aggttattac tgtg 24 <210> 1287 <211> 19 <212> DNA <213> CNAG_03059 diagnostic screening primer, pairing with B79 <400> 1287 aaagactcac ccttcaacg 19 <210> 1288 <211> 20 <212> DNA <213> CNAG_03059 Southern blot probe primer <400> 1288 cttttttccg ctggtttctc 20 <210> 1289 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 1289 ctttaaaggt ggtttgtg 18 <210> 1290 <211> 24 <212> DNA <213> STM common primer <400> 1290 gcatgccctg cccctaagaa ttcg 24 <210> 1291 <211> 21 <212> DNA <213> CNAG_00883 5 flanking region primer 1 <400> 1291 cgacagaaac aagtccaagt g 21 <210> 1292 <211> 36 <212> DNA <213> CNAG_00883 5 flanking region primer 2 <400> 1292 ctggccgtcg ttttacgtag tgaaatgagg attgcg 36 <210> 1293 <211> 35 <212> DNA <213> CNAG_00883 3 flanking region primer 1 <400> 1293 gtcatagctg tttcctgtac ctgcttttct tgcgg 35 <210> 1294 <211> 18 <212> DNA <213> CNAG_00883 3 flanking region primer 2 <400> 1294 acaagcccga atctcaag 18 <210> 1295 <211> 19 <212> DNA <213> CNAG_00883 diagnostic screening primer, pairing with B79 <400> 1295 cttttcactc aatgtcccg 19 <210> 1296 <211> 19 <212> DNA <213> CNAG_00883 Southern blot probe primer <400> 1296 taagaactgt tgccctgcc 19 <210> 1297 <211> 18 <212> DNA <213> CNAG_01551 5 flanking region primer 1 <400> 1297 tccgttgaga tagcgttg 18 <210> 1298 <211> 39 <212> DNA <213> CNAG_01551 5 flanking region primer 2 <400> 1298 tcactggccg tcgttttaca tggtggaggt gtaggactg 39 <210> 1299 <211> 39 <212> DNA <213> CNAG_01551 3 flanking region primer 1 <400> 1299 catggtcata gctgtttcct gttattcccc ctcaagagc 39 <210> 1300 <211> 20 <212> DNA <213> CNAG_01551 3 flanking region primer 2 <400> 1300 gcggcccatt cttcttgtcg 20 <210> 1301 <211> 18 <212> DNA <213> CNAG_01551 diagnostic screening primer, pairing with B79 <400> 1301 aaccttgcct aacaaccc 18 <210> 1302 <211> 20 <212> DNA <213> CNAG_01551 Southern blot probe primer <400> 1302 ggagtatggc tgaaatctgg 20 <210> 1303 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 1303 gagatctttc gggaggtctg gatt 24 <210> 1304 <211> 24 <212> DNA <213> STM common primer <400> 1304 gcatgccctg cccctaagaa ttcg 24 <210> 1305 <211> 20 <212> DNA <213> CNAG_04353 5 flanking region primer 1 <400> 1305 tgcgagagag aatgggagac 20 <210> 1306 <211> 38 <212> DNA <213> CNAG_04353 5 flanking region primer 2 <400> 1306 tcactggccg tcgttttacg gttccatagc gacatcag 38 <210> 1307 <211> 41 <212> DNA <213> CNAG_04353 3 flanking region primer 1 <400> 1307 catggtcata gctgtttcct gcgaaggttg tttgtgatac g 41 <210> 1308 <211> 20 <212> DNA <213> CNAG_04353 3 flanking region primer 2 <400> 1308 cccaacaact catcaatctc 20 <210> 1309 <211> 18 <212> DNA <213> CNAG_04353 diagnostic screening primer, pairing with B79 <400> 1309 gcattatcgt ttcccgac 18 <210> 1310 <211> 22 <212> DNA <213> CNAG_04353 Southern blot probe primer <400> 1310 ttgaggtagg tggagtgttg cc 22 <210> 1311 <211> 20 <212> DNA <213> CNAG_03346 5 flanking region primer 1 <400> 1311 cgccctttct attgttacac 20 <210> 1312 <211> 39 <212> DNA <213> CNAG_03346 5 flanking region primer 2 <400> 1312 tcactggccg tcgttttacg atgacaggag ggatgaatc 39 <210> 1313 <211> 43 <212> DNA <213> CNAG_03346 3 flanking region primer 1 <400> 1313 catggtcata gctgtttcct ggggagaata acgactcaat gtc 43 <210> 1314 <211> 18 <212> DNA <213> CNAG_03346 3 flanking region primer 2 <400> 1314 tcattgctga ctgggaag 18 <210> 1315 <211> 18 <212> DNA <213> CNAG_03346 diagnostic screening primer, pairing with B79 <400> 1315 aaagaggcgg tgttgaag 18 <210> 1316 <211> 19 <212> DNA <213> CNAG_03346 Southern blot probe primer <400> 1316 agccaggtaa tcttggagg 19 <210> 1317 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 1317 acacctacat caaaccctcc c 21 <210> 1318 <211> 24 <212> DNA <213> STM common primer <400> 1318 gcatgccctg cccctaagaa ttcg 24 <210> 1319 <211> 18 <212> DNA <213> CNAG_00239 5 flanking region primer 1 <400> 1319 aggattatcg cctgaacc 18 <210> 1320 <211> 37 <212> DNA <213> CNAG_00239 5 flanking region primer 2 <400> 1320 tcactggccg tcgttttaca tgttgttgaa gagggcg 37 <210> 1321 <211> 41 <212> DNA <213> CNAG_00239 3 flanking region primer 1 <400> 1321 catggtcata gctgtttcct gaacgacgac catcgcagta g 41 <210> 1322 <211> 18 <212> DNA <213> CNAG_00239 3 flanking region primer 2 <400> 1322 catcccgagt tggaaaac 18 <210> 1323 <211> 20 <212> DNA <213> CNAG_00239 diagnostic screening primer, pairing with B79 <400> 1323 cactggcgaa gtcaagtatg 20 <210> 1324 <211> 20 <212> DNA <213> CNAG_00239 Southern blot probe primer <400> 1324 ctactctgtg gtggcgtaag 20 <210> 1325 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 1325 cgcccgccct cactatccac 20 <210> 1326 <211> 24 <212> DNA <213> STM common primer <400> 1326 gcatgccctg cccctaagaa ttcg 24 <210> 1327 <211> 19 <212> DNA <213> CNAG_03129 5 flanking region primer 1 <400> 1327 gcaggcgaaa actattagg 19 <210> 1328 <211> 41 <212> DNA <213> CNAG_03129 5 flanking region primer 2 <400> 1328 gctcactggc cgtcgtttta cgtggggaag tcctcaaata c 41 <210> 1329 <211> 42 <212> DNA <213> CNAG_03129 3 flanking region primer 1 <400> 1329 catggtcata gctgtttcct gcgacgacat ctcaagtcag ag 42 <210> 1330 <211> 20 <212> DNA <213> CNAG_03129 3 flanking region primer 2 <400> 1330 tgatgggacg ataaaggttc 20 <210> 1331 <211> 19 <212> DNA <213> CNAG_03129 diagnostic screening primer, pairing with B79 <400> 1331 tgctttctct aatagcggc 19 <210> 1332 <211> 19 <212> DNA <213> CNAG_03129 Southern blot probe primer <400> 1332 gtgcttttga agttccgtc 19 <210> 1333 <211> 18 <212> DNA <213> CNAG_03826 5 flanking region primer 1 <400> 1333 aatgctgatt tggcaggg 18 <210> 1334 <211> 39 <212> DNA <213> CNAG_03826 5 flanking region primer 2 <400> 1334 tcactggccg tcgttttacc gatggaagat gttgtcagc 39 <210> 1335 <211> 39 <212> DNA <213> CNAG_03826 3 flanking region primer 1 <400> 1335 catggtcata gctgtttcct gacaaagcga aaggcagac 39 <210> 1336 <211> 18 <212> DNA <213> CNAG_03826 3 flanking region primer 2 <400> 1336 cgaaagtgat gcttcagc 18 <210> 1337 <211> 20 <212> DNA <213> CNAG_03826 diagnostic screening primer, pairing with B79 <400> 1337 ggaaggtcag gtcctatttg 20 <210> 1338 <211> 20 <212> DNA <213> CNAG_02671 5 flanking region primer 1 <400> 1338 cgccgagtta tctgacaaag 20 <210> 1339 <211> 41 <212> DNA <213> CNAG_02671 5 flanking region primer 2 <400> 1339 tcactggccg tcgttttact tctccaaaca ccgcccttga c 41 <210> 1340 <211> 40 <212> DNA <213> CNAG_02671 3 flanking region primer 1 <400> 1340 catggtcata gctgtttcct gtaacggggc ttagaagtgc 40 <210> 1341 <211> 18 <212> DNA <213> CNAG_02671 3 flanking region primer 2 <400> 1341 ttcatcaaag gtctggcg 18 <210> 1342 <211> 20 <212> DNA <213> CNAG_02671 diagnostic screening primer, pairing with B79 <400> 1342 cctgactcat cggcttactc 20 <210> 1343 <211> 18 <212> DNA <213> CNAG_02671 Southern blot probe primer <400> 1343 actcctgctt gcgttttc 18 <210> 1344 <211> 20 <212> DNA <213> CNAG_02134 5 flanking region primer 1 <400> 1344 ggggtaacta tcatcatccg 20 <210> 1345 <211> 38 <212> DNA <213> CNAG_02134 5 flanking region primer 2 <400> 1345 tcactggccg tcgttttact ctcgctcctt ttccttcc 38 <210> 1346 <211> 42 <212> DNA <213> CNAG_02134 3 flanking region primer 1 <400> 1346 catggtcata gctgtttcct ggcagtagaa ggtgatacaa cg 42 <210> 1347 <211> 19 <212> DNA <213> CNAG_02134 3 flanking region primer 2 <400> 1347 cgttacggtt ggacaatgg 19 <210> 1348 <211> 19 <212> DNA <213> CNAG_02134 diagnostic screening primer, pairing with B79 <400> 1348 cgtagaaaca acgcacgag 19 <210> 1349 <211> 21 <212> DNA <213> CNAG_02134 Southern blot probe primer <400> 1349 caagtattct ccagggttca g 21 <210> 1350 <211> 20 <212> DNA <213> CNAG_02936 5 flanking region primer 1 <400> 1350 gcgagtgatg acgagagttc 20 <210> 1351 <211> 39 <212> DNA <213> CNAG_02936 5 flanking region primer 2 <400> 1351 tcactggccg tcgttttacg ccttgaccgt ctcttctac 39 <210> 1352 <211> 42 <212> DNA <213> CNAG_02936 3 flanking region primer 1 <400> 1352 catggtcata gctgtttcct ggtctactcg tcactttcat cg 42 <210> 1353 <211> 21 <212> DNA <213> CNAG_02936 3 flanking region primer 2 <400> 1353 cgtcacaaca agcactctca c 21 <210> 1354 <211> 19 <212> DNA <213> CNAG_02936 diagnostic screening primer, pairing with B79 <400> 1354 ggatgttcag gtgccaatc 19 <210> 1355 <211> 18 <212> DNA <213> CNAG_07460 5 flanking region primer 1 <400> 1355 tcggcattgg catacttg 18 <210> 1356 <211> 39 <212> DNA <213> CNAG_07460 5 flanking region primer 2 <400> 1356 gctcactggc cgtcgtttta cgtgatttga gcggtgaag 39 <210> 1357 <211> 42 <212> DNA <213> CNAG_07460 3 flanking region primer 1 <400> 1357 catggtcata gctgtttcct gccaaagtgg actcaaatag gg 42 <210> 1358 <211> 22 <212> DNA <213> CNAG_07460 3 flanking region primer 2 <400> 1358 ccgttacttt ccaaatcagg ag 22 <210> 1359 <211> 20 <212> DNA <213> CNAG_07460 diagnostic screening primer, pairing with B79 <400> 1359 cgtctgaacc tgattctgag 20 <210> 1360 <211> 19 <212> DNA <213> CNAG_07460 Southern blot probe primer <400> 1360 ccgatgacca aaagtaggc 19 <210> 1361 <211> 19 <212> DNA <213> CNAG_07460 Southern blot probe primer <400> 1361 cagagttgat tgccgaaag 19 <210> 1362 <211> 19 <212> DNA <213> CNAG_05436 5 flanking region primer 1 <400> 1362 gcccttttgc tttaccttc 19 <210> 1363 <211> 41 <212> DNA <213> CNAG_05436 5 flanking region primer 2 <400> 1363 tcactggccg tcgttttacc gtagagggat gattttgaga c 41 <210> 1364 <211> 40 <212> DNA <213> CNAG_05436 3 flanking region primer 1 <400> 1364 catggtcata gctgtttcct ggcatccttc gtgctaacag 40 <210> 1365 <211> 18 <212> DNA <213> CNAG_05436 3 flanking region primer 2 <400> 1365 gccttgaaga acaaacgg 18 <210> 1366 <211> 19 <212> DNA <213> CNAG_05436 diagnostic screening primer, pairing with B79 <400> 1366 gggataggag gcattgatg 19 <210> 1367 <211> 20 <212> DNA <213> CNAG_05436 Southern blot probe primer <400> 1367 caccaaagtg gatttccttg 20 <210> 1368 <211> 21 <212> DNA <213> CNAG_05436 Southern blot probe primer <400> 1368 ctcggacaaa tcactctact g 21 <210> 1369 <211> 21 <212> DNA <213> CNAG_02698 5 flanking region primer 1 <400> 1369 ctacagataa tgtgctcagc c 21 <210> 1370 <211> 37 <212> DNA <213> CNAG_02698 5 flanking region primer 2 <400> 1370 tcactggccg tcgttttact ttgttccata gccagcg 37 <210> 1371 <211> 40 <212> DNA <213> CNAG_02698 3 flanking region primer 1 <400> 1371 catggtcata gctgtttcct ggttctgtct tgaagggtgc 40 <210> 1372 <211> 20 <212> DNA <213> CNAG_02698 3 flanking region primer 2 <400> 1372 cggtgaagtt ggctaagaac 20 <210> 1373 <211> 19 <212> DNA <213> CNAG_02698 diagnostic screening primer, pairing with B79 <400> 1373 gaaagggcta agaagcgac 19 <210> 1374 <211> 20 <212> DNA <213> CNAG_02698 Southern blot probe primer <400> 1374 aaactctcca tctgtgtccc 20 <210> 1375 <211> 20 <212> DNA <213> CNAG_04600 5 flanking region primer 1 <400> 1375 gctgttggct gattgtttac 20 <210> 1376 <211> 39 <212> DNA <213> CNAG_04600 5 flanking region primer 2 <400> 1376 tcactggccg tcgttttact tacagtatcc caccgtccc 39 <210> 1377 <211> 40 <212> DNA <213> CNAG_04600 3 flanking region primer 1 <400> 1377 catggtcata gctgtttcct gtgggagtga tgtctttggc 40 <210> 1378 <211> 18 <212> DNA <213> CNAG_04600 3 flanking region primer 2 <400> 1378 ccctcaaaag ttgcgaag 18 <210> 1379 <211> 18 <212> DNA <213> CNAG_04600 diagnostic screening primer, pairing with B79 <400> 1379 ctggattttg gaatgggc 18 <210> 1380 <211> 18 <212> DNA <213> CNAG_04600 Southern blot probe primer <400> 1380 ttacatttgc cagcggac 18 <110> AmtixBio Co., Ltd. <120> Novel genes for regulating the virulence of Cryptococcus neoformans and their use <130> AMTIX16P-0001.2-KR <160> 1380 <170> KopatentIn 2.0 <210> 1 <211> 19 <212> DNA <213> CNAG_03132 5 flanking region primer 1 <400> 1 gcgtggcatt tagagagtg 19 <210> 2 <211> 38 <212> DNA <213> CNAG_03132 5 flanking region primer 2 <400> 2 tcactggccg tcgttttacg agcgtattgg atgaggag 38 <210> 3 <211> 39 <212> DNA <213> CNAG_03132 3 flanking region primer 1 <400> 3 catggtcata gctgtttcct gaaacctcgc tttgagacg 39 <210> 4 <211> 18 <212> DNA <213> CNAG_03132 3 flanking region primer 2 <400> 4 tgacgaagcg acagaatc 18 <210> 5 <211> 20 <212> DNA <213> CNAG_03132 diagnostic screening primer, pairing with B79 <400> 5 cccaaacact attatcgcag 20 <210> 6 <211> 18 <212> DNA <213> CNAG_03132 Southern blot probe primer <400> 6 tactcgtatg aatcggcg 18 <210> 7 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 7 tgctagaggg cgggagagtt 20 <210> 8 <211> 24 <212> DNA <213> STM common primer <400> 8 gcatgccctg cccctaagaa ttcg 24 <210> 9 <211> 20 <212> DNA <213> CNAG_05380 5 flanking region primer 1 <400> 9 cctactgctt ggcacattac 20 <210> 10 <211> 35 <212> DNA <213> CNAG_05380 5 flanking region primer 2 <400> 10 ctggccgtcg ttttaccgtc gtttcttttt gggag 35 <210> 11 <211> 37 <212> DNA <213> CNAG_05380 3 flanking region primer 1 <400> 11 gtcatagctg tttcctgtcc atctccagtc tgtgaac 37 <210> 12 <211> 20 <212> DNA <213> CNAG_05380 3 flanking region primer 2 <400> 12 cgcttcttag gacctcactc 20 <210> 13 <211> 20 <212> DNA <213> CNAG_05380 diagnostic screening primer, pairing with B79 <400> 13 tctgaaggcg gaatctggac 20 <210> 14 <211> 24 <212> DNA <213> CNAG_05380 Southern blot probe primer <400> 14 ggagcaagga actaaagatg gtag 24 <210> 15 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 15 tgctagaggg cgggagagtt 20 <210> 16 <211> 24 <212> DNA <213> STM common primer <400> 16 gcatgccctg cccctaagaa ttcg 24 <210> 17 <211> 20 <212> DNA <213> CNAG_06719 5 flanking region primer 1 <400> 17 tcgccaactg aacaaacccc 20 <210> 18 <211> 36 <212> DNA <213> CNAG_06719 5 flanking region primer 2 <400> 18 ctggccgtcg ttttactccc gcctaacact ttcgtg 36 <210> 19 <211> 36 <212> DNA <213> CNAG_06719 3 flanking region primer 1 <400> 19 gtcatagctg tttcctgtgt caagccataa agagcc 36 <210> 20 <211> 20 <212> DNA <213> CNAG_06719 3 flanking region primer 2 <400> 20 ctgatagttt tgccacttgc 20 <210> 21 <211> 22 <212> DNA <213> CNAG_06719 diagnostic screening primer, pairing with B79 <400> 21 aataccctcc ttacttcccc gc 22 <210> 22 <211> 18 <212> DNA <213> CNAG_06719 Southern blot probe primer <400> 22 gtaaaagttt gggcgtgg 18 <210> 23 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 23 tgctagaggg cgggagagtt 20 <210> 24 <211> 24 <212> DNA <213> STM common primer <400> 24 gcatgccctg cccctaagaa ttcg 24 <210> 25 <211> 20 <212> DNA <213> CNAG_04093 5 flanking region primer 1 <400> 25 tccaggcgta ttcatcccag 20 <210> 26 <211> 38 <212> DNA <213> CNAG_04093 5 flanking region primer 2 <400> 26 ctggccgtcg ttttacaagg attgtgacct caggctcg 38 <210> 27 <211> 36 <212> DNA <213> CNAG_04093 3 flanking region primer 1 <400> 27 gtcatagctg tttcctgatg tttctcctcc agagcc 36 <210> 28 <211> 19 <212> DNA <213> CNAG_04093 3 flanking region primer 2 <400> 28 gcaagaacca aatgacagc 19 <210> 29 <211> 24 <212> DNA <213> CNAG_04093 diagnostic screening primer, pairing with B79 <400> 29 atcattcctc tgacaagtcg ggcg 24 <210> 30 <211> 19 <212> DNA <213> CNAG_04093 Southern blot probe primer <400> 30 atgtgtcctg gatggcttg 19 <210> 31 <211> 20 <212> DNA <213> NAT#5 STM primer <400> 31 tgctagaggg cgggagagtt 20 <210> 32 <211> 24 <212> DNA <213> STM common primer <400> 32 gcatgccctg cccctaagaa ttcg 24 <210> 33 <211> 18 <212> DNA <213> CNAG_04518 5 flanking region primer 1 <400> 33 tgtcctttct tgtgaccg 18 <210> 34 <211> 39 <212> DNA <213> CNAG_04518 5 flanking region primer 2 <400> 34 tcactggccg tcgttttacc agtaacgaag aacgcaatc 39 <210> 35 <211> 40 <212> DNA <213> CNAG_04518 3 flanking region primer 1 <400> 35 catggtcata gctgtttcct gcacaagcca cgattattgc 40 <210> 36 <211> 20 <212> DNA <213> CNAG_04518 3 flanking region primer 2 <400> 36 tgtatctgtg tgtggactgc 20 <210> 37 <211> 18 <212> DNA <213> CNAG_04518 diagnostic screening primer, pairing with B79 <400> 37 acacaaaggg aggaatgc 18 <210> 38 <211> 19 <212> DNA <213> CNAG_04518 Southern blot probe primer <400> 38 catacactgt gccacgaac 19 <210> 39 <211> 20 <212> DNA <213> NAT#6 STM primer <400> 39 atagctacca cacgatagct 20 <210> 40 <211> 24 <212> DNA <213> STM common primer <400> 40 gcatgccctg cccctaagaa ttcg 24 <210> 41 <211> 20 <212> DNA <213> CNAG_01858 5 flanking region primer 1 <400> 41 tcttcaggaa cataggatgg 20 <210> 42 <211> 38 <212> DNA <213> CNAG_01858 5 flanking region primer 2 <400> 42 tcactggccg tcgttttacg agattgttgg gaggattg 38 <210> 43 <211> 39 <212> DNA <213> CNAG_01858 3 flanking region primer 1 <400> 43 catggtcata gctgtttcct ggggcgaata aggtttctg 39 <210> 44 <211> 18 <212> DNA <213> CNAG_01858 3 flanking region primer 2 <400> 44 tgttttcgct catccgtc 18 <210> 45 <211> 19 <212> DNA <213> CNAG_01858 diagnostic screening primer, pairing with B79 <400> 45 aacacagcgg aatgacctc 19 <210> 46 <211> 18 <212> DNA <213> CNAG_01858 Southern blot probe primer <400> 46 cacttggttc attgctgg 18 <210> 47 <211> 20 <212> DNA <213> NAT#43 STM primer <400> 47 ccagctacca atcacgctac 20 <210> 48 <211> 24 <212> DNA <213> STM common primer <400> 48 gcatgccctg cccctaagaa ttcg 24 <210> 49 <211> 22 <212> DNA <213> CNAG_02305 5 flanking region primer 1 <400> 49 ccagaaggat gtgatactga gg 22 <210> 50 <211> 36 <212> DNA <213> CNAG_02305 5 flanking region primer 2 <400> 50 ctggccgtcg ttttacgaaa tcgttctgaa actgcg 36 <210> 51 <211> 39 <212> DNA <213> CNAG_02305 3 flanking region primer 1 <400> 51 gtcatagctg tttcctgatc gttcacaggc tcaaggctg 39 <210> 52 <211> 22 <212> DNA <213> CNAG_02305 3 flanking region primer 2 <400> 52 gaggtcatta gagggtattt gc 22 <210> 53 <211> 19 <212> DNA <213> CNAG_02305 diagnostic screening primer, pairing with B79 <400> 53 gcgtggcgaa tacatacag 19 <210> 54 <211> 19 <212> DNA <213> CNAG_02305 Southern blot probe primer <400> 54 aggtgctaca aagactcgg 19 <210> 55 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 55 cgcaaaatca ctagccctat agcg 24 <210> 56 <211> 24 <212> DNA <213> STM common primer <400> 56 gcatgccctg cccctaagaa ttcg 24 <210> 57 <211> 19 <212> DNA <213> CNAG_06156 5 flanking region primer 1 <400> 57 aagatagcac ctccgacag 19 <210> 58 <211> 41 <212> DNA <213> CNAG_06156 5 flanking region primer 2 <400> 58 gctcactggc cgtcgtttta cgctgttgcg aaagataaag g 41 <210> 59 <211> 41 <212> DNA <213> CNAG_06156 3 flanking region primer 1 <400> 59 catggtcata gctgtttcct gccaggaatg aactaccaag g 41 <210> 60 <211> 19 <212> DNA <213> CNAG_06156 3 flanking region primer 2 <400> 60 ttgaagcagg actctccac 19 <210> 61 <211> 19 <212> DNA <213> CNAG_06156 diagnostic screening primer, pairing with B79 <400> 61 tgcttgtcgt ttggttagg 19 <210> 62 <211> 19 <212> DNA <213> CNAG_06156 Southern blot probe primer <400> 62 ggtgaaaatg acggtccag 19 <210> 63 <211> 24 <212> DNA <213> NAT#119 STM primer <400> 63 ctccccacat aaagagagct aaac 24 <210> 64 <211> 24 <212> DNA <213> STM common primer <400> 64 gcatgccctg cccctaagaa ttcg 24 <210> 65 <211> 21 <212> DNA <213> CNAG_06188 5 flanking region primer 1 <400> 65 ctcttgtctt ctgcctattc c 21 <210> 66 <211> 35 <212> DNA <213> CNAG_06188 5 flanking region primer 2 <400> 66 ctggccgtcg ttttacggac atagtgaaac tgggg 35 <210> 67 <211> 38 <212> DNA <213> CNAG_06188 3 flanking region primer 1 <400> 67 gtcatagctg tttcctgtca tcctccgact ctaaactg 38 <210> 68 <211> 20 <212> DNA <213> CNAG_06188 3 flanking region primer 2 <400> 68 gttttcctac ttttgtgggc 20 <210> 69 <211> 20 <212> DNA <213> CNAG_06188 diagnostic screening primer, pairing with B79 <400> 69 agaagacgac tgtggtaggc 20 <210> 70 <211> 20 <212> DNA <213> CNAG_06188 Southern blot probe primer <400> 70 gagtcagcag cagaagattc 20 <210> 71 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 71 acagctccaa acctcgctaa acag 24 <210> 72 <211> 24 <212> DNA <213> STM common primer <400> 72 gcatgccctg cccctaagaa ttcg 24 <210> 73 <211> 19 <212> DNA <213> CNAG_05785 5 flanking region primer 1 <400> 73 catttctact tggttgccg 19 <210> 74 <211> 36 <212> DNA <213> CNAG_05785 5 flanking region primer 2 <400> 74 ctggccgtcg ttttacagga gaaagtgatg gtcggc 36 <210> 75 <211> 39 <212> DNA <213> CNAG_05785 3 flanking region primer 1 <400> 75 gtcatagctg tttcctgtta tcgtgagcct tcaagcccc 39 <210> 76 <211> 21 <212> DNA <213> CNAG_05785 3 flanking region primer 2 <400> 76 ccattgtaga tgtcggttac g 21 <210> 77 <211> 22 <212> DNA <213> CNAG_05785 diagnostic screening primer, pairing with B79 <400> 77 ttgcggtctt tgtcccttgt gg 22 <210> 78 <211> 24 <212> DNA <213> CNAG_05785 Southern blot probe primer <400> 78 ttctccactt gctcgcctat cctc 24 <210> 79 <211> 24 <212> DNA <213> NAT#125 STM primer <400> 79 cgctacagcc agcgcgcgca agcg 24 <210> 80 <211> 24 <212> DNA <213> STM common primer <400> 80 gcatgccctg cccctaagaa ttcg 24 <210> 81 <211> 20 <212> DNA <213> CNAG_05019 5 flanking region primer 1 <400> 81 aaccaccaca atgtcttcag 20 <210> 82 <211> 40 <212> DNA <213> CNAG_05019 5 flanking region primer 2 <400> 82 gctcactggc cgtcgtttta cagacccttg tttggttctg 40 <210> 83 <211> 41 <212> DNA <213> CNAG_05019 3 flanking region primer 1 <400> 83 catggtcata gctgtttcct gagtggagag ggagagcaat g 41 <210> 84 <211> 18 <212> DNA <213> CNAG_05019 3 flanking region primer 2 <400> 84 tttacgaaga gcagcgac 18 <210> 85 <211> 18 <212> DNA <213> CNAG_05019 diagnostic screening primer, pairing with B79 <400> 85 agatagcaac ggctttgg 18 <210> 86 <211> 20 <212> DNA <213> CNAG_05019 Southern blot probe primer <400> 86 cgcaaaaggg aagatggata 20 <210> 87 <211> 20 <212> DNA <213> NAT#150 STM primer <400> 87 acatacaccc ccatcccccc 20 <210> 88 <211> 24 <212> DNA <213> STM common primer <400> 88 gcatgccctg cccctaagaa ttcg 24 <210> 89 <211> 18 <212> DNA <213> CNAG_02877 5 flanking region primer 1 <400> 89 gtttaccatt tcaccgcc 18 <210> 90 <211> 39 <212> DNA <213> CNAG_02877 5 flanking region primer 2 <400> 90 tcactggccg tcgttttact gatgaggata cattcggag 39 <210> 91 <211> 41 <212> DNA <213> CNAG_02877 3 flanking region primer 1 <400> 91 catggtcata gctgtttcct gggcttcaac aatcaccatt c 41 <210> 92 <211> 18 <212> DNA <213> CNAG_02877 3 flanking region primer 2 <400> 92 acgaaaagcg gatagcag 18 <210> 93 <211> 20 <212> DNA <213> CNAG_02877 diagnostic screening primer, pairing with B79 <400> 93 agccttatta tgaccaaccc 20 <210> 94 <211> 20 <212> DNA <213> CNAG_02877 Southern blot probe primer <400> 94 caagttcctt tgcctacacc 20 <210> 95 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 95 acgcaccaga cacacaacca g 21 <210> 96 <211> 24 <212> DNA <213> STM common primer <400> 96 gcatgccctg cccctaagaa ttcg 24 <210> 97 <211> 20 <212> DNA <213> CNAG_00039 5 flanking region primer 1 <400> 97 tcactctttc ggggcatcag 20 <210> 98 <211> 38 <212> DNA <213> CNAG_00039 5 flanking region primer 2 <400> 98 ctggccgtcg ttttacctaa ctggggatga aatgtctg 38 <210> 99 <211> 39 <212> DNA <213> CNAG_00039 3 flanking region primer 1 <400> 99 gtcatagctg tttcctgggg gaagagcaga aagagtaac 39 <210> 100 <211> 22 <212> DNA <213> CNAG_00039 3 flanking region primer 2 <400> 100 gcaatcgtaa acaacaccct tg 22 <210> 101 <211> 20 <212> DNA <213> CNAG_00039 diagnostic screening primer, pairing with B79 <400> 101 cgtctgtcgt ttatcggttg 20 <210> 102 <211> 19 <212> DNA <213> CNAG_00039 Southern blot probe primer <400> 102 accctgtaac atttgtccg 19 <210> 103 <211> 19 <212> DNA <213> CNAG_00039 Southern blot probe primer <400> 103 acttaccctt cacgccaag 19 <210> 104 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 104 caccaactcc ccatctccat 20 <210> 105 <211> 24 <212> DNA <213> STM common primer <400> 105 gcatgccctg cccctaagaa ttcg 24 <210> 106 <211> 20 <212> DNA <213> CNAG_00039 5 flanking region primer 1 <400> 106 agacattggg cttgacggtg 20 <210> 107 <211> 39 <212> DNA <213> CNAG_00039 5 flanking region primer 2 <400> 107 tcactggccg tcgttttaca cgagttggag aggggttac 39 <210> 108 <211> 39 <212> DNA <213> CNAG_00039 3 flanking region primer 1 <400> 108 catggtcata gctgtttcct gtagaaagga atcggggac 39 <210> 109 <211> 20 <212> DNA <213> CNAG_00039 3 flanking region primer 2 <400> 109 ggagaacgaa aacaaagtgc 20 <210> 110 <211> 19 <212> DNA <213> CNAG_00039 diagnostic screening primer, pairing with B79 <400> 110 ggttcttgtt tatgccctg 19 <210> 111 <211> 20 <212> DNA <213> CNAG_00039 Southern blot probe primer <400> 111 aacgagggta gattgcgtcg 20 <210> 112 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 112 caccaactcc ccatctccat 20 <210> 113 <211> 24 <212> DNA <213> STM common primer <400> 113 gcatgccctg cccctaagaa ttcg 24 <210> 114 <211> 19 <212> DNA <213> CNAG_01014 5 flanking region primer 1 <400> 114 tatcgccgac atctctcag 19 <210> 115 <211> 38 <212> DNA <213> CNAG_01014 5 flanking region primer 2 <400> 115 tcactggccg tcgttttact ttaggacgga ggggattg 38 <210> 116 <211> 39 <212> DNA <213> CNAG_01014 3 flanking region primer 1 <400> 116 catggtcata gctgtttcct ggagccgaat caggaaatc 39 <210> 117 <211> 19 <212> DNA <213> CNAG_01014 3 flanking region primer 2 <400> 117 cgcaggcttt tctactctc 19 <210> 118 <211> 30 <212> DNA <213> CNAG_01014 diagnostic screening primer, pairing with B79 <400> 118 cgcgcggccg cgaaaacctc cacagagcag 30 <210> 119 <211> 19 <212> DNA <213> CNAG_01014 Southern blot probe primer <400> 119 ttgcttgctg tctcgtaac 19 <210> 120 <211> 18 <212> DNA <213> CNAG_01014 Southern blot probe primer <400> 120 gatttcctga ttcggctc 18 <210> 121 <211> 20 <212> DNA <213> CTAGAGCCCGCCACAACGCT <400> 121 ctagagcccg ccacaacgct 20 <210> 122 <211> 24 <212> DNA <213> STM common primer <400> 122 gcatgccctg cccctaagaa ttcg 24 <210> 123 <211> 21 <212> DNA <213> CNAG_06762 5 flanking region primer 1 <400> 123 tgggtgagag aaagatgtca g 21 <210> 124 <211> 39 <212> DNA <213> CNAG_06762 5 flanking region primer 2 <400> 124 tcactggccg tcgttttact gtgagtggtt gtgctgtgg 39 <210> 125 <211> 41 <212> DNA <213> CNAG_06762 3 flanking region primer 1 <400> 125 catggtcata gctgtttcct gggctcatag gtgttgtaac g 41 <210> 126 <211> 20 <212> DNA <213> CNAG_06762 3 flanking region primer 2 <400> 126 agtattcggt ggtgacaaag 20 <210> 127 <211> 21 <212> DNA <213> CNAG_06762 diagnostic screening primer, pairing with B79 <400> 127 tgggtgagag aaagatgtca g 21 <210> 128 <211> 19 <212> DNA <213> CNAG_06762 Southern blot probe primer <400> 128 tgttccaccc cttgctaac 19 <210> 129 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 129 ctccacatcc atcgctccaa 20 <210> 130 <211> 24 <212> DNA <213> STM common primer <400> 130 gcatgccctg cccctaagaa ttcg 24 <210> 131 <211> 18 <212> DNA <213> CNAG_04630 5 flanking region primer 1 <400> 131 atggaaggac tttgacgg 18 <210> 132 <211> 40 <212> DNA <213> CNAG_04630 5 flanking region primer 2 <400> 132 tcactggccg tcgttttact gactcacact tgattctccc 40 <210> 133 <211> 40 <212> DNA <213> CNAG_04630 3 flanking region primer 1 <400> 133 gatggtcata gctgtttcct gctcattttc cctttcgtcg 40 <210> 134 <211> 18 <212> DNA <213> CNAG_04630 3 flanking region primer 2 <400> 134 taaaggaatg cggctgag 18 <210> 135 <211> 19 <212> DNA <213> CNAG_04630 diagnostic screening primer, pairing with B79 <400> 135 ccaacagcaa cttcgtttc 19 <210> 136 <211> 21 <212> DNA <213> CNAG_04630 Southern blot probe primer <400> 136 tctcttctcc ttggaactga g 21 <210> 137 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 137 ctccacatcc atcgctccaa 20 <210> 138 <211> 24 <212> DNA <213> STM common primer <400> 138 gcatgccctg cccctaagaa ttcg 24 <210> 139 <211> 19 <212> DNA <213> CNAG_03401 5 flanking region primer 1 <400> 139 ttcctcgtcc aagtaaacc 19 <210> 140 <211> 41 <212> DNA <213> CNAG_03401 5 flanking region primer 2 <400> 140 gctcactggc cgtcgtttta caatctcgca aaccgcacac g 41 <210> 141 <211> 43 <212> DNA <213> CNAG_03401 3 flanking region primer 1 <400> 141 catggtcata gctgtttcct gagcgcgaac agggaaatac acc 43 <210> 142 <211> 18 <212> DNA <213> CNAG_03401 3 flanking region primer 2 <400> 142 tcgtgtagga atgggtag 18 <210> 143 <211> 20 <212> DNA <213> CNAG_03401 diagnostic screening primer, pairing with B79 <400> 143 tggtctcccc gacgatttac 20 <210> 144 <211> 19 <212> DNA <213> CNAG_03401 Southern blot probe primer <400> 144 tatggtcagg aggaagcag 19 <210> 145 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 145 cagatctcga acgataccca 20 <210> 146 <211> 24 <212> DNA <213> STM common primer <400> 146 gcatgccctg cccctaagaa ttcg 24 <210> 147 <211> 22 <212> DNA <213> CNAG_04594 5 flanking region primer 1 <400> 147 tggcatcgtg ctgttacaag tc 22 <210> 148 <211> 45 <212> DNA <213> CNAG_04594 5 flanking region primer 2 <400> 148 gctcactggc cgtcgtttta catttgcgga tggacctgcc ttagg 45 <210> 149 <211> 41 <212> DNA <213> CNAG_04594 3 flanking region primer 1 <400> 149 catggtcata gctgtttcct gatcaggatt caggacacga c 41 <210> 150 <211> 18 <212> DNA <213> CNAG_04594 3 flanking region primer 2 <400> 150 tttggttctc tctcccac 18 <210> 151 <211> 19 <212> DNA <213> CNAG_04594 diagnostic screening primer, pairing with B79 <400> 151 attgcttgag accagcgtc 19 <210> 152 <211> 18 <212> DNA <213> CNAG_04594 Southern blot probe primer <400> 152 acggttgctt tgctatgc 18 <210> 153 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 153 cagatctcga acgataccca 20 <210> 154 <211> 24 <212> DNA <213> STM common primer <400> 154 gcatgccctg cccctaagaa ttcg 24 <210> 155 <211> 19 <212> DNA <213> CNAG_07901 5 flanking region primer 1 <400> 155 actcaccttc ccgctttag 19 <210> 156 <211> 34 <212> DNA <213> CNAG_07901 5 flanking region primer 2 <400> 156 ctggccgtcg ttttacgttt gagttgaggt tgcg 34 <210> 157 <211> 36 <212> DNA <213> CNAG_07901 3 flanking region primer 1 <400> 157 gtcatagctg tttcctgaga atgttgatgg tagcgg 36 <210> 158 <211> 19 <212> DNA <213> CNAG_07901 3 flanking region primer 2 <400> 158 tccaccgaag aaactgaag 19 <210> 159 <211> 18 <212> DNA <213> CNAG_07901 diagnostic screening primer, pairing with B79 <400> 159 gggatttgga tacgcttg 18 <210> 160 <211> 18 <212> DNA <213> CNAG_07901 Southern blot probe primer <400> 160 ttgcggtggg aaagtatc 18 <210> 161 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 161 ccatagaact agctaaagca 20 <210> 162 <211> 24 <212> DNA <213> STM common primer <400> 162 gcatgccctg cccctaagaa ttcg 24 <210> 163 <211> 19 <212> DNA <213> CNAG_04588 5 flanking region primer 1 <400> 163 aaaagaggtc cgatgagtg 19 <210> 164 <211> 40 <212> DNA <213> CNAG_04588 5 flanking region primer 2 <400> 164 gctcactggc cgtcgtttta cgtgaatgat aaaggcgacc 40 <210> 165 <211> 39 <212> DNA <213> CNAG_04588 3 flanking region primer 1 <400> 165 catggtcata gctgtttcct gtgctgtttc actatccgc 39 <210> 166 <211> 19 <212> DNA <213> CNAG_04588 3 flanking region primer 2 <400> 166 cgaaagcgac tggagtaac 19 <210> 167 <211> 20 <212> DNA <213> CNAG_04588 diagnostic screening primer, pairing with B79 <400> 167 gaagatgctg aagcgtattc 20 <210> 168 <211> 18 <212> DNA <213> CNAG_04588 Southern blot probe primer <400> 168 acacttatcc gccattcc 18 <210> 169 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 169 ccatagaact agctaaagca 20 <210> 170 <211> 24 <212> DNA <213> STM common primer <400> 170 gcatgccctg cccctaagaa ttcg 24 <210> 171 <211> 18 <212> DNA <213> CNAG_03998 5 flanking region primer 1 <400> 171 gccattgtga aagcatag 18 <210> 172 <211> 43 <212> DNA <213> CNAG_03998 5 flanking region primer 2 <400> 172 gctcactggc cgtcgtttta cacctgtcta tctcactgtc cag 43 <210> 173 <211> 39 <212> DNA <213> CNAG_03998 3 flanking region primer 1 <400> 173 catggtcata gctgtttcct gtgttcagca gcaaaaggc 39 <210> 174 <211> 18 <212> DNA <213> CNAG_03998 3 flanking region primer 2 <400> 174 gcggagagaa ggattatc 18 <210> 175 <211> 20 <212> DNA <213> CNAG_03998 diagnostic screening primer, pairing with B79 <400> 175 gctgacaact tggaatcatc 20 <210> 176 <211> 20 <212> DNA <213> CNAG_03998 Southern blot probe primer <400> 176 gggcttctct tcttcttcac 20 <210> 177 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 177 ccagccagca ctccgatata 20 <210> 178 <211> 24 <212> DNA <213> STM common primer <400> 178 gcatgccctg cccctaagaa ttcg 24 <210> 179 <211> 20 <212> DNA <213> CNAG_04804 5 flanking region primer 1 <400> 179 taggtagttc ctgctcgtgg 20 <210> 180 <211> 40 <212> DNA <213> CNAG_04804 5 flanking region primer 2 <400> 180 tcactggccg tcgttttacc atcttgtcct gtaatgaggc 40 <210> 181 <211> 39 <212> DNA <213> CNAG_04804 3 flanking region primer 1 <400> 181 catggtcata gctgtttcct ggattggagg gcgatttag 39 <210> 182 <211> 20 <212> DNA <213> CNAG_04804 3 flanking region primer 2 <400> 182 caacaaggtc caagaaactg 20 <210> 183 <211> 21 <212> DNA <213> CNAG_04804 diagnostic screening primer, pairing with B79 <400> 183 agaggactga gacttttgtc g 21 <210> 184 <211> 19 <212> DNA <213> CNAG_04804 Southern blot probe primer <400> 184 cgaaggggaa ctggttatc 19 <210> 185 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 185 ggtgttggat cggggtggat 20 <210> 186 <211> 24 <212> DNA <213> STM common primer <400> 186 gcatgccctg cccctaagaa ttcg 24 <210> 187 <211> 20 <212> DNA <213> CNAG_06134 5 flanking region primer 1 <400> 187 gtttgaggct ggtaaaaagg 20 <210> 188 <211> 39 <212> DNA <213> CNAG_06134 5 flanking region primer 2 <400> 188 gctcactggc cgtcgtttta catggggaat gaaagcgtg 39 <210> 189 <211> 41 <212> DNA <213> CNAG_06134 3 flanking region primer 1 <400> 189 catggtcata gctgtttcct gaaggggcga gagtagttca g 41 <210> 190 <211> 21 <212> DNA <213> CNAG_06134 3 flanking region primer 2 <400> 190 gactgtaaag gagggcataa g 21 <210> 191 <211> 19 <212> DNA <213> CNAG_06134 diagnostic screening primer, pairing with B79 <400> 191 aactcttctc agccttcgg 19 <210> 192 <211> 19 <212> DNA <213> CNAG_06134 Southern blot probe primer <400> 192 cgttctccgt cttgatagc 19 <210> 193 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 193 acacctacat caaaccctcc c 21 <210> 194 <211> 24 <212> DNA <213> STM common primer <400> 194 gcatgccctg cccctaagaa ttcg 24 <210> 195 <211> 19 <212> DNA <213> CNAG_04798 5 flanking region primer 1 <400> 195 caatgattag gcttgcgtc 19 <210> 196 <211> 38 <212> DNA <213> CNAG_04798 5 flanking region primer 2 <400> 196 tcactggccg tcgttttact gttgttgggc agtggtag 38 <210> 197 <211> 39 <212> DNA <213> CNAG_04798 3 flanking region primer 1 <400> 197 catggtcata gctgtttcct gtttagggaa gaggaagcg 39 <210> 198 <211> 18 <212> DNA <213> CNAG_04798 3 flanking region primer 2 <400> 198 tgtcatccag cagattgg 18 <210> 199 <211> 19 <212> DNA <213> CNAG_04798 diagnostic screening primer, pairing with B79 <400> 199 tgaatgactg atggacccc 19 <210> 200 <211> 18 <212> DNA <213> CNAG_04798 Southern blot probe primer <400> 200 tcggttctcg ttttggag 18 <210> 201 <211> 20 <212> DNA <213> CNAG_04864 5 flanking region primer 1 <400> 201 tcatttcttc gtcctctgag 20 <210> 202 <211> 36 <212> DNA <213> CNAG_04864 5 flanking region primer 2 <400> 202 ctggccgtcg ttttacgctc cccataatca gactcc 36 <210> 203 <211> 36 <212> DNA <213> CNAG_04864 3 flanking region primer 1 <400> 203 gtcatagctg tttcctgacg aatggagttt ctggag 36 <210> 204 <211> 20 <212> DNA <213> CNAG_04864 3 flanking region primer 2 <400> 204 gcgttctcca tagttgtagc 20 <210> 205 <211> 20 <212> DNA <213> CNAG_04864 diagnostic screening primer, pairing with B79 <400> 205 atgtagataa agcacgcacc 20 <210> 206 <211> 20 <212> DNA <213> CNAG_04864 Southern blot probe primer <400> 206 cgagttattg taggttgggc 20 <210> 207 <211> 20 <212> DNA <213> CNAG_01973 5 flanking region primer 1 <400> 207 gcttgaggtt gaaacaggtc 20 <210> 208 <211> 41 <212> DNA <213> CNAG_01973 5 flanking region primer 2 <400> 208 tcactggccg tcgttttacc agatgtgtgt aagagggtca g 41 <210> 209 <211> 39 <212> DNA <213> CNAG_01973 3 flanking region primer 1 <400> 209 catggtcata gctgtttcct gtaccgagaa cgatttggc 39 <210> 210 <211> 20 <212> DNA <213> CNAG_01973 3 flanking region primer 2 <400> 210 tcccataaga ccacacactg 20 <210> 211 <211> 19 <212> DNA <213> CNAG_01973 diagnostic screening primer, pairing with B79 <400> 211 tgatgctgaa tagacgggg 19 <210> 212 <211> 19 <212> DNA <213> CNAG_01973 Southern blot probe primer <400> 212 agccttgacg acatacctg 19 <210> 213 <211> 20 <212> DNA <213> NAT#6 STM primer <400> 213 atagctacca cacgatagct 20 <210> 214 <211> 24 <212> DNA <213> STM common primer <400> 214 gcatgccctg cccctaagaa ttcg 24 <210> 215 <211> 19 <212> DNA <213> CNAG_03914 5 flanking region primer 1 <400> 215 gatgtcgttg ttccagagc 19 <210> 216 <211> 35 <212> DNA <213> CNAG_03914 5 flanking region primer 2 <400> 216 ctggccgtcg ttttactttg gatgtaagag gtcgg 35 <210> 217 <211> 37 <212> DNA <213> CNAG_03914 3 flanking region primer 1 <400> 217 gtcatagctg tttcctgcga gtatccttac cgtgtcg 37 <210> 218 <211> 19 <212> DNA <213> CNAG_03914 3 flanking region primer 2 <400> 218 acactcctgc ccaagaaac 19 <210> 219 <211> 19 <212> DNA <213> CNAG_03914 diagnostic screening primer, pairing with B79 <400> 219 actccactcc atccaaagc 19 <210> 220 <211> 20 <212> DNA <213> CNAG_03914 Southern blot probe primer <400> 220 ataagaggtc cttttgctcc 20 <210> 221 <211> 20 <212> DNA <213> NAT#43 STM primer <400> 221 ccagctacca atcacgctac 20 <210> 222 <211> 24 <212> DNA <213> STM common primer <400> 222 gcatgccctg cccctaagaa ttcg 24 <210> 223 <211> 19 <212> DNA <213> CNAG_00828 5 flanking region primer 1 <400> 223 accagtggca aggttagag 19 <210> 224 <211> 39 <212> DNA <213> CNAG_00828 5 flanking region primer 2 <400> 224 tcactggccg tcgttttact ggcagaagca caagattag 39 <210> 225 <211> 40 <212> DNA <213> CNAG_00828 3 flanking region primer 1 <400> 225 catggtcata gctgtttcct ggccatttct cgtagcgaac 40 <210> 226 <211> 19 <212> DNA <213> CNAG_00828 3 flanking region primer 2 <400> 226 actgcttatc cccacatcg 19 <210> 227 <211> 20 <212> DNA <213> CNAG_00828 diagnostic screening primer, pairing with B79 <400> 227 tgtgaagtgt gatgatgtcc 20 <210> 228 <211> 19 <212> DNA <213> CNAG_00828 Southern blot probe primer <400> 228 ggttgccact gataaaagc 19 <210> 229 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 229 cgcaaaatca ctagccctat agcg 24 <210> 230 <211> 24 <212> DNA <213> STM common primer <400> 230 gcatgccctg cccctaagaa ttcg 24 <210> 231 <211> 19 <212> DNA <213> CNAG_00871 5 flanking region primer 1 <400> 231 atagcgggca agtgtctac 19 <210> 232 <211> 38 <212> DNA <213> CNAG_00871 5 flanking region primer 2 <400> 232 tcactggccg tcgttttacc caacctttca ttccagtc 38 <210> 233 <211> 42 <212> DNA <213> CNAG_00871 3 flanking region primer 1 <400> 233 catggtcata gctgtttcct gggatacaaa tctcaacccc tg 42 <210> 234 <211> 19 <212> DNA <213> CNAG_00871 3 flanking region primer 2 <400> 234 ggttttcact tcaagagcg 19 <210> 235 <211> 18 <212> DNA <213> CNAG_00871 diagnostic screening primer, pairing with B79 <400> 235 tccaatagca ccacaacc 18 <210> 236 <211> 19 <212> DNA <213> CNAG_00871 Southern blot probe primer <400> 236 aacatccctt tactcggcg 19 <210> 237 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 237 ccatagcgat atctacccca atct 24 <210> 238 <211> 24 <212> DNA <213> STM common primer <400> 238 gcatgccctg cccctaagaa ttcg 24 <210> 239 <211> 19 <212> DNA <213> CNAG_07464 5 flanking region primer 1 <400> 239 ggcatcagga tagaaacgc 19 <210> 240 <211> 41 <212> DNA <213> CNAG_07464 5 flanking region primer 2 <400> 240 gctcactggc cgtcgtttta cggggtgaga tttgaaggta g 41 <210> 241 <211> 40 <212> DNA <213> CNAG_07464 3 flanking region primer 1 <400> 241 catggtcata gctgtttcct ggatgtgact tggtcttggg 40 <210> 242 <211> 20 <212> DNA <213> CNAG_07464 3 flanking region primer 2 <400> 242 tggtccagtc tcctcattac 20 <210> 243 <211> 18 <212> DNA <213> CNAG_07464 diagnostic screening primer, pairing with B79 <400> 243 tcttccatct cagcatcg 18 <210> 244 <211> 24 <212> DNA <213> CNAG_07464 Southern blot probe primer <400> 244 ccatagcgat atctacccca atct 24 <210> 245 <211> 20 <212> DNA <213> NAT#150 STM primer <400> 245 acatacaccc ccatcccccc 20 <210> 246 <211> 24 <212> DNA <213> STM common primer <400> 246 gcatgccctg cccctaagaa ttcg 24 <210> 247 <211> 23 <212> DNA <213> CNAG_00460 5 flanking region primer 1 <400> 247 cgtgggagag tgagtaaagt atg 23 <210> 248 <211> 42 <212> DNA <213> CNAG_00460 5 flanking region primer 2 <400> 248 gctcactggc cgtcgtttta ccgggtaaga gtggatgtat gg 42 <210> 249 <211> 39 <212> DNA <213> CNAG_00460 3 flanking region primer 1 <400> 249 catggtcata gctgtttcct gaagggcacc gttcattag 39 <210> 250 <211> 19 <212> DNA <213> CNAG_00460 3 flanking region primer 2 <400> 250 ttcttaggat gcgggactc 19 <210> 251 <211> 18 <212> DNA <213> CNAG_00460 diagnostic screening primer, pairing with B79 <400> 251 tgcgttgaca aagtcgtg 18 <210> 252 <211> 22 <212> DNA <213> CNAG_00460 Southern blot probe primer <400> 252 tagactccag taaggtccaa tc 22 <210> 253 <211> 24 <212> DNA <213> NAT#213 STM primer <400> 253 ctggggattt tgatgtgtct atgt 24 <210> 254 <211> 24 <212> DNA <213> STM common primer <400> 254 gcatgccctg cccctaagaa ttcg 24 <210> 255 <211> 18 <212> DNA <213> CNAG_01431 5 flanking region primer 1 <400> 255 tccatttggg ggtttcag 18 <210> 256 <211> 39 <212> DNA <213> CNAG_01431 5 flanking region primer 2 <400> 256 tcactggccg tcgttttact gtcttcgttc ttgccctcc 39 <210> 257 <211> 42 <212> DNA <213> CNAG_01431 3 flanking region primer 1 <400> 257 catggtcata gctgtttcct gcgttgacag aagaggacaa gg 42 <210> 258 <211> 20 <212> DNA <213> CNAG_01431 3 flanking region primer 2 <400> 258 tgacagatga tgagcagagg 20 <210> 259 <211> 19 <212> DNA <213> CNAG_01431 diagnostic screening primer, pairing with B79 <400> 259 attccagagc ctcacttgc 19 <210> 260 <211> 21 <212> DNA <213> CNAG_01431 Southern blot probe primer <400> 260 cctgcttgtc actacactga g 21 <210> 261 <211> 24 <212> DNA <213> NAT#213 STM primer <400> 261 ctggggattt tgatgtgtct atgt 24 <210> 262 <211> 24 <212> DNA <213> STM common primer <400> 262 gcatgccctg cccctaagaa ttcg 24 <210> 263 <211> 21 <212> DNA <213> CNAG_03894 5 flanking region primer 1 <400> 263 catctatctc aacggggtat c 21 <210> 264 <211> 39 <212> DNA <213> CNAG_03894 5 flanking region primer 2 <400> 264 gctcactggc cgtcgtttta ctttttcggg agaacgcag 39 <210> 265 <211> 40 <212> DNA <213> CNAG_03894 3 flanking region primer 1 <400> 265 catggtcata gctgtttcct gaggaccaag aatgctgagg 40 <210> 266 <211> 22 <212> DNA <213> CNAG_03894 3 flanking region primer 2 <400> 266 ttctcccttg tcgccgataa gc 22 <210> 267 <211> 20 <212> DNA <213> CNAG_03894 diagnostic screening primer, pairing with B79 <400> 267 tcttatcgct caccacatcc 20 <210> 268 <211> 19 <212> DNA <213> CNAG_03894 Southern blot probe primer <400> 268 ggaacattgg gaaaaggtg 19 <210> 269 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 269 cagatctcga acgataccca 20 <210> 270 <211> 24 <212> DNA <213> STM common primer <400> 270 gcatgccctg cccctaagaa ttcg 24 <210> 271 <211> 18 <212> DNA <213> CNAG_06483 5 flanking region primer 1 <400> 271 tggagcagat ggagtttg 18 <210> 272 <211> 35 <212> DNA <213> CNAG_06483 5 flanking region primer 2 <400> 272 ctggccgtcg ttttacgctt ctccctattt ccagc 35 <210> 273 <211> 36 <212> DNA <213> CNAG_06483 3 flanking region primer 1 <400> 273 gtcatagctg tttcctgaat ggagcccaga gaacag 36 <210> 274 <211> 19 <212> DNA <213> CNAG_06483 3 flanking region primer 2 <400> 274 tctcgctgag ggaaaactc 19 <210> 275 <211> 18 <212> DNA <213> CNAG_06483 diagnostic screening primer, pairing with B79 <400> 275 ccctttctct gcgttttg 18 <210> 276 <211> 18 <212> DNA <213> CNAG_06483 Southern blot probe primer <400> 276 caatgaagaa ctggtgcg 18 <210> 277 <211> 20 <212> DNA <213> NAT#227 STM primer <400> 277 tcgtggttta gagggagcgc 20 <210> 278 <211> 24 <212> DNA <213> STM common primer <400> 278 gcatgccctg cccctaagaa ttcg 24 <210> 279 <211> 18 <212> DNA <213> CNAG_00896 5 flanking region primer 1 <400> 279 aaagggtcac gatggaac 18 <210> 280 <211> 35 <212> DNA <213> CNAG_00896 5 flanking region primer 2 <400> 280 ctggccgtcg ttttacgcga ttctttactc ctggc 35 <210> 281 <211> 36 <212> DNA <213> CNAG_00896 3 flanking region primer 1 <400> 281 gtcatagctg tttcctgttc tgagacctcc gtaacc 36 <210> 282 <211> 19 <212> DNA <213> CNAG_00896 3 flanking region primer 2 <400> 282 ctgtatcgct ttagccacc 19 <210> 283 <211> 19 <212> DNA <213> CNAG_00896 diagnostic screening primer, pairing with B79 <400> 283 tacaaggacc tcacgaacg 19 <210> 284 <211> 20 <212> DNA <213> CNAG_00896 Southern blot probe primer <400> 284 acgagcagag ggacagatag 20 <210> 285 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 285 gagagatccc aacatcacgc 20 <210> 286 <211> 24 <212> DNA <213> STM common primer <400> 286 gcatgccctg cccctaagaa ttcg 24 <210> 287 <211> 18 <212> DNA <213> CNAG_01977 5 flanking region primer 1 <400> 287 tgggaaaacg aagcactc 18 <210> 288 <211> 35 <212> DNA <213> CNAG_01977 5 flanking region primer 2 <400> 288 ctggccgtcg ttttactagg aggtcacttt cgctc 35 <210> 289 <211> 36 <212> DNA <213> CNAG_01977 3 flanking region primer 1 <400> 289 gtcatagctg tttcctgcgg caaagaacaa ggtatg 36 <210> 290 <211> 20 <212> DNA <213> CNAG_01977 3 flanking region primer 2 <400> 290 tgatgggctt gaacgaacgc 20 <210> 291 <211> 19 <212> DNA <213> CNAG_01977 diagnostic screening primer, pairing with B79 <400> 291 tgtcggcaca catctactg 19 <210> 292 <211> 22 <212> DNA <213> CNAG_01977 Southern blot probe primer <400> 292 gtctcagcct cttgttacag at 22 <210> 293 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 293 gagagatccc aacatcacgc 20 <210> 294 <211> 24 <212> DNA <213> STM common primer <400> 294 gcatgccctg cccctaagaa ttcg 24 <210> 295 <211> 19 <212> DNA <213> CNAG_01069 5 flanking region primer 1 <400> 295 cttttatcta tggcgagcg 19 <210> 296 <211> 39 <212> DNA <213> CNAG_01069 5 flanking region primer 2 <400> 296 gctcactggc cgtcgtttta ctatcgcact gctggaatc 39 <210> 297 <211> 40 <212> DNA <213> CNAG_01069 3 flanking region primer 1 <400> 297 catggtcata gctgtttcct gctactttcc ctcacatccg 40 <210> 298 <211> 19 <212> DNA <213> CNAG_01069 3 flanking region primer 2 <400> 298 tggagggtag atgcgttac 19 <210> 299 <211> 20 <212> DNA <213> CNAG_01069 diagnostic screening primer, pairing with B79 <400> 299 agagaagcgg aagagaaaac 20 <210> 300 <211> 20 <212> DNA <213> CNAG_01069 Southern blot probe primer <400> 300 ccttgcgaaa atgtcttctc 20 <210> 301 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 301 atttcgatgt atatatgtag c 21 <210> 302 <211> 24 <212> DNA <213> STM common primer <400> 302 gcatgccctg cccctaagaa ttcg 24 <210> 303 <211> 19 <212> DNA <213> CNAG_06276 5 flanking region primer 1 <400> 303 tcaagtctac gcagtcgtg 19 <210> 304 <211> 40 <212> DNA <213> CNAG_06276 5 flanking region primer 2 <400> 304 gctcactggc cgtcgtttta cgcagcaatc aactcactcg 40 <210> 305 <211> 41 <212> DNA <213> CNAG_06276 3 flanking region primer 1 <400> 305 catggtcata gctgtttcct gaggagagtg ggagtatgga g 41 <210> 306 <211> 18 <212> DNA <213> CNAG_06276 3 flanking region primer 2 <400> 306 acccatcaaa aggcgtag 18 <210> 307 <211> 20 <212> DNA <213> CNAG_06276 diagnostic screening primer, pairing with B79 <400> 307 accatttgtc aactgtcagg 20 <210> 308 <211> 20 <212> DNA <213> CNAG_06276 Southern blot probe primer <400> 308 tcgctcactg acaagaagac 20 <210> 309 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 309 atttcgatgt atatatgtag c 21 <210> 310 <211> 24 <212> DNA <213> STM common primer <400> 310 gcatgccctg cccctaagaa ttcg 24 <210> 311 <211> 19 <212> DNA <213> CNAG_02476 5 flanking region primer 1 <400> 311 tctgctcaat gaaacgggc 19 <210> 312 <211> 45 <212> DNA <213> CNAG_02476 5 flanking region primer 2 <400> 312 gctcactggc cgtcgtttta cgcctttgct tcctttgctt cttac 45 <210> 313 <211> 40 <212> DNA <213> CNAG_02476 3 flanking region primer 1 <400> 313 catggtcata gctgtttcct ggtgtcttga atccaatgcc 40 <210> 314 <211> 20 <212> DNA <213> CNAG_02476 3 flanking region primer 2 <400> 314 ctgctccata caaggcttag 20 <210> 315 <211> 20 <212> DNA <213> CNAG_02476 diagnostic screening primer, pairing with B79 <400> 315 tcgtcagcaa ttgaaaacct 20 <210> 316 <211> 19 <212> DNA <213> CNAG_02476 Southern blot probe primer <400> 316 atcaagtcag accacgagg 19 <210> 317 <211> 20 <212> DNA <213> NAT#219 STM primer <400> 317 ccctaaaacc ctacagcaat 20 <210> 318 <211> 24 <212> DNA <213> STM common primer <400> 318 gcatgccctg cccctaagaa ttcg 24 <210> 319 <211> 20 <212> DNA <213> CNAG_01841 5 flanking region primer 1 <400> 319 ccattatcat ccgtcaccac 20 <210> 320 <211> 37 <212> DNA <213> CNAG_01841 5 flanking region primer 2 <400> 320 ctggccgtcg ttttacgaag agggaagaga gggtaac 37 <210> 321 <211> 36 <212> DNA <213> CNAG_01841 3 flanking region primer 1 <400> 321 gtcatagctg tttcctgtga ggctgaggaa acagag 36 <210> 322 <211> 18 <212> DNA <213> CNAG_01841 3 flanking region primer 2 <400> 322 tccttttccc aacctgtc 18 <210> 323 <211> 21 <212> DNA <213> CNAG_01841 diagnostic screening primer, pairing with B79 <400> 323 ttcagtggtt ggtctctaca g 21 <210> 324 <211> 21 <212> DNA <213> CNAG_01841 Southern blot probe primer <400> 324 tggtaaaggt ctatcagtgc c 21 <210> 325 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 325 atgtaggtag ggtgataggt 20 <210> 326 <211> 24 <212> DNA <213> STM common primer <400> 326 gcatgccctg cccctaagaa ttcg 24 <210> 327 <211> 19 <212> DNA <213> CNAG_04457 5 flanking region primer 1 <400> 327 cttcaaaatc cagagcagg 19 <210> 328 <211> 38 <212> DNA <213> CNAG_04457 5 flanking region primer 2 <400> 328 ctggccgtcg ttttacctgg tctgatagtt cgtattcg 38 <210> 329 <211> 39 <212> DNA <213> CNAG_04457 3 flanking region primer 1 <400> 329 gtcatagctg tttcctggga cttgaacaat ggtatggac 39 <210> 330 <211> 19 <212> DNA <213> CNAG_04457 3 flanking region primer 2 <400> 330 tttagagcct acggtggtc 19 <210> 331 <211> 22 <212> DNA <213> CNAG_04457 diagnostic screening primer, pairing with B79 <400> 331 ttgcccgacg agatgctatg ac 22 <210> 332 <211> 20 <212> DNA <213> CNAG_04457 Southern blot probe primer <400> 332 tccgtctccg atgatttgcg 20 <210> 333 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 333 atgtaggtag ggtgataggt 20 <210> 334 <211> 24 <212> DNA <213> STM common primer <400> 334 gcatgccctg cccctaagaa ttcg 24 <210> 335 <211> 19 <212> DNA <213> CNAG_06252 5 flanking region primer 1 <400> 335 tctgacaggt tcaatcgcc 19 <210> 336 <211> 41 <212> DNA <213> CNAG_06252 5 flanking region primer 2 <400> 336 tcactggccg tcgttttacg catttctaca ccgtgattag c 41 <210> 337 <211> 40 <212> DNA <213> CNAG_06252 3 flanking region primer 1 <400> 337 catggtcata gctgtttcct gggatgtgcg gttttgttag 40 <210> 338 <211> 19 <212> DNA <213> CNAG_06252 3 flanking region primer 2 <400> 338 taacgaagtt ctccccgtg 19 <210> 339 <211> 20 <212> DNA <213> CNAG_06252 diagnostic screening primer, pairing with B79 <400> 339 tgagagggaa cgaaggatac 20 <210> 340 <211> 20 <212> DNA <213> CNAG_06252 Southern blot probe primer <400> 340 cttcgcctgt tggaaataag 20 <210> 341 <211> 20 <212> DNA <213> CNAG_06327 5 flanking region primer 1 <400> 341 ggagaaggga aagatgggtg 20 <210> 342 <211> 41 <212> DNA <213> CNAG_06327 5 flanking region primer 2 <400> 342 gctcactggc cgtcgtttta ctgtttccac tattgggaga g 41 <210> 343 <211> 40 <212> DNA <213> CNAG_06327 3 flanking region primer 1 <400> 343 catggtcata gctgtttcct gaggtgaagg tgtgagattg 40 <210> 344 <211> 18 <212> DNA <213> CNAG_06327 3 flanking region primer 2 <400> 344 caataatgtc ctcgccag 18 <210> 345 <211> 19 <212> DNA <213> CNAG_06327 diagnostic screening primer, pairing with B79 <400> 345 gtatgaaggg aaggtgctg 19 <210> 346 <211> 18 <212> DNA <213> CNAG_06327 Southern blot probe primer <400> 346 acgggttacg agaaatgc 18 <210> 347 <211> 19 <212> DNA <213> CNAG_03790 5 flanking region primer 1 <400> 347 tgagggttgt tgagggaag 19 <210> 348 <211> 40 <212> DNA <213> CNAG_03790 5 flanking region primer 2 <400> 348 gctcactggc cgtcgtttta cagcactgta tcgccgaaac 40 <210> 349 <211> 42 <212> DNA <213> CNAG_03790 3 flanking region primer 1 <400> 349 catggtcata gctgtttcct gctcgtagca atgtagatgt gc 42 <210> 350 <211> 21 <212> DNA <213> CNAG_03790 3 flanking region primer 2 <400> 350 tcagatgaag acgactacga c 21 <210> 351 <211> 19 <212> DNA <213> CNAG_03790 diagnostic screening primer, pairing with B79 <400> 351 cttccacaaa gccagattc 19 <210> 352 <211> 18 <212> DNA <213> CNAG_03790 Southern blot probe primer <400> 352 agattgcgtg aaacccac 18 <210> 353 <211> 19 <212> DNA <213> CNAG_05112 5 flanking region primer 1 <400> 353 atgtgcgaag tccaagagc 19 <210> 354 <211> 40 <212> DNA <213> CNAG_05112 5 flanking region primer 2 <400> 354 gctcactggc cgtcgtttta cgggtgaaga caacagatgg 40 <210> 355 <211> 41 <212> DNA <213> CNAG_05112 3 flanking region primer 1 <400> 355 catggtcata gctgtttcct gcagaccaac tggcgtaata g 41 <210> 356 <211> 19 <212> DNA <213> CNAG_05112 3 flanking region primer 2 <400> 356 acgaccaaat gctcttcgc 19 <210> 357 <211> 19 <212> DNA <213> CNAG_05112 diagnostic screening primer, pairing with B79 <400> 357 tcggatacag gcgatagag 19 <210> 358 <211> 19 <212> DNA <213> CNAG_05112 Southern blot probe primer <400> 358 tcaacagcag tttcaggtg 19 <210> 359 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 359 acagctccaa acctcgctaa acag 24 <210> 360 <211> 24 <212> DNA <213> STM common primer <400> 360 gcatgccctg cccctaagaa ttcg 24 <210> 361 <211> 19 <212> DNA <213> CNAG_00068 5 flanking region primer 1 <400> 361 tctatcacct ctgcgttgc 19 <210> 362 <211> 39 <212> DNA <213> CNAG_00068 5 flanking region primer 2 <400> 362 tcactggccg tcgttttaca ttcctaccac tatcccctg 39 <210> 363 <211> 40 <212> DNA <213> CNAG_00068 3 flanking region primer 1 <400> 363 catggtcata gctgtttcct gcggataaga tgcctgagac 40 <210> 364 <211> 22 <212> DNA <213> CNAG_00068 3 flanking region primer 2 <400> 364 ggtatgtttc tcggatgtaa gg 22 <210> 365 <211> 20 <212> DNA <213> CNAG_00068 diagnostic screening primer, pairing with B79 <400> 365 catcatcagg tatcagcagc 20 <210> 366 <211> 20 <212> DNA <213> CNAG_00068 Southern blot probe primer <400> 366 cttccaactt caatgttccc 20 <210> 367 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 367 cgcaaaatca ctagccctat agcg 24 <210> 368 <211> 24 <212> DNA <213> STM common primer <400> 368 gcatgccctg cccctaagaa ttcg 24 <210> 369 <211> 19 <212> DNA <213> CNAG_04184 5 flanking region primer 1 <400> 369 tcgttcacat aactgccac 19 <210> 370 <211> 38 <212> DNA <213> CNAG_04184 5 flanking region primer 2 <400> 370 tcactggccg tcgttttact cattgtgagc ctcgtatg 38 <210> 371 <211> 39 <212> DNA <213> CNAG_04184 3 flanking region primer 1 <400> 371 catggtcata gctgtttcct gaatggtgac ggttcaagg 39 <210> 372 <211> 18 <212> DNA <213> CNAG_04184 3 flanking region primer 2 <400> 372 cttttccact gaatccgc 18 <210> 373 <211> 19 <212> DNA <213> CNAG_04184 diagnostic screening primer, pairing with B79 <400> 373 tggtccgatg ccgtaaatc 19 <210> 374 <211> 19 <212> DNA <213> CNAG_04184 Southern blot probe primer <400> 374 caagggaatg actcactcc 19 <210> 375 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 375 ccatagcgat atctacccca atct 24 <210> 376 <211> 24 <212> DNA <213> STM common primer <400> 376 gcatgccctg cccctaagaa ttcg 24 <210> 377 <211> 18 <212> DNA <213> CNAG_03018 5 flanking region primer 1 <400> 377 agccgtttgt cattctgc 18 <210> 378 <211> 37 <212> DNA <213> CNAG_03018 5 flanking region primer 2 <400> 378 tcactggccg tcgttttacg tcgttgctca ttttgcg 37 <210> 379 <211> 39 <212> DNA <213> CNAG_03018 3 flanking region primer 1 <400> 379 catggtcata gctgtttcct ggcgaacctc caataaagc 39 <210> 380 <211> 18 <212> DNA <213> CNAG_03018 3 flanking region primer 2 <400> 380 cccttttttc cctgcttc 18 <210> 381 <211> 18 <212> DNA <213> CNAG_03018 diagnostic screening primer, pairing with B79 <400> 381 aatagggatg ccaaccag 18 <210> 382 <211> 18 <212> DNA <213> CNAG_03018 Southern blot probe primer <400> 382 caaaaagtgt cagcacgc 18 <210> 383 <211> 20 <212> DNA <213> NAT#150 STM primer <400> 383 acatacaccc ccatcccccc 20 <210> 384 <211> 24 <212> DNA <213> STM common primer <400> 384 gcatgccctg cccctaagaa ttcg 24 <210> 385 <211> 20 <212> DNA <213> CNAG_01708 5 flanking region primer 1 <400> 385 ccgactcgtt cgtctatctg 20 <210> 386 <211> 41 <212> DNA <213> CNAG_01708 5 flanking region primer 2 <400> 386 tcactggccg tcgttttact ggagaggata gatgtgatga c 41 <210> 387 <211> 41 <212> DNA <213> CNAG_01708 3 flanking region primer 1 <400> 387 catggtcata gctgtttcct gaacttctac ccaggtcgtg c 41 <210> 388 <211> 18 <212> DNA <213> CNAG_01708 3 flanking region primer 2 <400> 388 aactgccctt cttcaagc 18 <210> 389 <211> 20 <212> DNA <213> CNAG_01708 diagnostic screening primer, pairing with B79 <400> 389 tagcctctcc caatgtccac 20 <210> 390 <211> 20 <212> DNA <213> CNAG_01708 Southern blot probe primer <400> 390 cgattctcct ttgctgagtc 20 <210> 391 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 391 acgcaccaga cacacaacca g 21 <210> 392 <211> 24 <212> DNA <213> STM common primer <400> 392 gcatgccctg cccctaagaa ttcg 24 <210> 393 <211> 19 <212> DNA <213> CNAG_05861 5 flanking region primer 1 <400> 393 ggttttggtc acttcaccg 19 <210> 394 <211> 38 <212> DNA <213> CNAG_05861 5 flanking region primer 2 <400> 394 tcactggccg tcgttttaca gcagaaagga gatgatgc 38 <210> 395 <211> 40 <212> DNA <213> CNAG_05861 3 flanking region primer 1 <400> 395 catggtcata gctgtttcct ggacgacttt tacgaccagg 40 <210> 396 <211> 18 <212> DNA <213> CNAG_05861 3 flanking region primer 2 <400> 396 aaatggaaca agcccgtc 18 <210> 397 <211> 18 <212> DNA <213> CNAG_05861 diagnostic screening primer, pairing with B79 <400> 397 tctgaagggc gtaaatgc 18 <210> 398 <211> 18 <212> DNA <213> CNAG_05861 Southern blot probe primer <400> 398 aaggaatggg ttgttggg 18 <210> 399 <211> 24 <212> DNA <213> NAT#184 STM primer <400> 399 atatatggct cgagctagat agag 24 <210> 400 <211> 24 <212> DNA <213> STM common primer <400> 400 gcatgccctg cccctaagaa ttcg 24 <210> 401 <211> 20 <212> DNA <213> CNAG_02066 5 flanking region primer 1 <400> 401 ccccattgta aacattggtg 20 <210> 402 <211> 36 <212> DNA <213> CNAG_02066 5 flanking region primer 2 <400> 402 ctggccgtcg ttttaccgtg atgtggagtt gattcg 36 <210> 403 <211> 36 <212> DNA <213> CNAG_02066 3 flanking region primer 1 <400> 403 gtcatagctg tttcctgtgc tgccaatgga aggttc 36 <210> 404 <211> 20 <212> DNA <213> CNAG_02066 3 flanking region primer 2 <400> 404 atagtctcgc tccttggtgc 20 <210> 405 <211> 19 <212> DNA <213> CNAG_02066 diagnostic screening primer, pairing with B79 <400> 405 gttcgttatc cgctctttg 19 <210> 406 <211> 19 <212> DNA <213> CNAG_02066 Southern blot probe primer <400> 406 acataggcga gacagttgc 19 <210> 407 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 407 atatggatgt ttttagcgag 20 <210> 408 <211> 24 <212> DNA <213> STM common primer <400> 408 gcatgccctg cccctaagaa ttcg 24 <210> 409 <211> 19 <212> DNA <213> CNAG_03527 5 flanking region primer 1 <400> 409 ccatctcacg gttgatacg 19 <210> 410 <211> 35 <212> DNA <213> CNAG_03527 5 flanking region primer 2 <400> 410 ctggccgtcg ttttactcaa acccttatcc caccg 35 <210> 411 <211> 37 <212> DNA <213> CNAG_03527 3 flanking region primer 1 <400> 411 gtcatagctg tttcctgtct ttccctgctt tgcctgc 37 <210> 412 <211> 20 <212> DNA <213> CNAG_03527 3 flanking region primer 2 <400> 412 cgtcatccgt cgtgtaactg 20 <210> 413 <211> 19 <212> DNA <213> CNAG_03527 diagnostic screening primer, pairing with B79 <400> 413 tctcgcaagg tcagagttc 19 <210> 414 <211> 21 <212> DNA <213> CNAG_03527 Southern blot probe primer <400> 414 ctatgagcga cattcacaga c 21 <210> 415 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 415 gatctctcgc gcttggggga 20 <210> 416 <211> 24 <212> DNA <213> STM common primer <400> 416 gcatgccctg cccctaagaa ttcg 24 <210> 417 <211> 18 <212> DNA <213> CNAG_04263 5 flanking region primer 1 <400> 417 tgtttccaca caccgatg 18 <210> 418 <211> 36 <212> DNA <213> CNAG_04263 5 flanking region primer 2 <400> 418 ctggccgtcg ttttacggtg gtggtcatct gtatgc 36 <210> 419 <211> 37 <212> DNA <213> CNAG_04263 3 flanking region primer 1 <400> 419 gtcatagctg tttcctgaga agatggagac gcacacg 37 <210> 420 <211> 19 <212> DNA <213> CNAG_04263 3 flanking region primer 2 <400> 420 gctattgcgt gaagactcc 19 <210> 421 <211> 20 <212> DNA <213> CNAG_04263 diagnostic screening primer, pairing with B79 <400> 421 cgtgattgag agtagcaacc 20 <210> 422 <211> 19 <212> DNA <213> CNAG_04263 Southern blot probe primer <400> 422 gtaatgtcag tgagtgcgg 19 <210> 423 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 423 tatccccctc tccgctctct agca 24 <210> 424 <211> 24 <212> DNA <213> STM common primer <400> 424 gcatgccctg cccctaagaa ttcg 24 <210> 425 <211> 20 <212> DNA <213> CNAG_00018 5 flanking region primer 1 <400> 425 tttgcctcaa tctctctcag 20 <210> 426 <211> 41 <212> DNA <213> CNAG_00018 5 flanking region primer 2 <400> 426 gctcactggc cgtcgtttta cggaatacaa catccacctc c 41 <210> 427 <211> 42 <212> DNA <213> CNAG_00018 3 flanking region primer 1 <400> 427 catggtcata gctgtttcct ggcacgggtt acattctaca tc 42 <210> 428 <211> 20 <212> DNA <213> CNAG_00018 3 flanking region primer 2 <400> 428 ggggaggact tgaacttgag 20 <210> 429 <211> 18 <212> DNA <213> CNAG_00018 diagnostic screening primer, pairing with B79 <400> 429 aacaaactgc cttcctcc 18 <210> 430 <211> 19 <212> DNA <213> CNAG_00018 Southern blot probe primer <400> 430 gcttggtcgg ataaagatg 19 <210> 431 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 431 gcggtcgctt tatagcgatt 20 <210> 432 <211> 24 <212> DNA <213> STM common primer <400> 432 gcatgccctg cccctaagaa ttcg 24 <210> 433 <211> 19 <212> DNA <213> CNAG_05176 5 flanking region primer 1 <400> 433 tgagaaggag gatggtcac 19 <210> 434 <211> 37 <212> DNA <213> CNAG_05176 5 flanking region primer 2 <400> 434 tcactggccg tcgttttaca tgtggatagt gttgcgg 37 <210> 435 <211> 40 <212> DNA <213> CNAG_05176 3 flanking region primer 1 <400> 435 catggtcata gctgtttcct ggcaaaaagg gagaatgtcg 40 <210> 436 <211> 19 <212> DNA <213> CNAG_05176 3 flanking region primer 2 <400> 436 tcatacagcc gagttgacc 19 <210> 437 <211> 19 <212> DNA <213> CNAG_05176 diagnostic screening primer, pairing with B79 <400> 437 tgtgagtatg cctatcccg 19 <210> 438 <211> 19 <212> DNA <213> CNAG_05176 Southern blot probe primer <400> 438 tcaccgagac taatggcac 19 <210> 439 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 439 gcggtcgctt tatagcgatt 20 <210> 440 <211> 24 <212> DNA <213> STM common primer <400> 440 gcatgccctg cccctaagaa ttcg 24 <210> 441 <211> 19 <212> DNA <213> CNAG_06339 5 flanking region primer 1 <400> 441 tagtgaaaga ctcggcacc 19 <210> 442 <211> 37 <212> DNA <213> CNAG_06339 5 flanking region primer 2 <400> 442 tcactggccg tcgttttacc agtcaccgca aagattc 37 <210> 443 <211> 41 <212> DNA <213> CNAG_06339 3 flanking region primer 1 <400> 443 catggtcata gctgtttcct gcgcttcgtt attacctgtt c 41 <210> 444 <211> 22 <212> DNA <213> CNAG_06339 3 flanking region primer 2 <400> 444 ctccaggata agtagtgtga gg 22 <210> 445 <211> 20 <212> DNA <213> CNAG_06339 diagnostic screening primer, pairing with B79 <400> 445 gggaaagtag ggtaaccacc 20 <210> 446 <211> 20 <212> DNA <213> CNAG_06339 Southern blot probe primer <400> 446 gaggagaggt gaataatgcc 20 <210> 447 <211> 24 <212> DNA <213> NAT#213 STM primer <400> 447 ctggggattt tgatgtgtct atgt 24 <210> 448 <211> 24 <212> DNA <213> STM common primer <400> 448 gcatgccctg cccctaagaa ttcg 24 <210> 449 <211> 19 <212> DNA <213> CNAG_07593 5 flanking region primer 1 <400> 449 gattgcttgg atggacttc 19 <210> 450 <211> 39 <212> DNA <213> CNAG_07593 5 flanking region primer 2 <400> 450 tcactggccg tcgttttact gcctttcttg ccgaactcg 39 <210> 451 <211> 40 <212> DNA <213> CNAG_07593 3 flanking region primer 1 <400> 451 catggtcata gctgtttcct gtttgaagga ccaggagcag 40 <210> 452 <211> 19 <212> DNA <213> CNAG_07593 3 flanking region primer 2 <400> 452 tcaagtgaca cagtgccac 19 <210> 453 <211> 19 <212> DNA <213> CNAG_07593 diagnostic screening primer, pairing with B79 <400> 453 ccataatcag acagcgttg 19 <210> 454 <211> 19 <212> DNA <213> CNAG_07593 Southern blot probe primer <400> 454 ggtttggggg aatagtgag 19 <210> 455 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 455 ctccacatcc atcgctccaa 20 <210> 456 <211> 24 <212> DNA <213> STM common primer <400> 456 gcatgccctg cccctaagaa ttcg 24 <210> 457 <211> 20 <212> DNA <213> CNAG_00670 5 flanking region primer 1 <400> 457 tgatgaagat gctgatgagg 20 <210> 458 <211> 41 <212> DNA <213> CNAG_00670 5 flanking region primer 2 <400> 458 gctcactggc cgtcgtttta ctcatttggc gtctcactta c 41 <210> 459 <211> 39 <212> DNA <213> CNAG_00670 3 flanking region primer 1 <400> 459 catggtcata gctgtttcct gtgtgaaagg ggagtgatg 39 <210> 460 <211> 19 <212> DNA <213> CNAG_00670 3 flanking region primer 2 <400> 460 agaatacatc gtggttggc 19 <210> 461 <211> 22 <212> DNA <213> CNAG_00670 diagnostic screening primer, pairing with B79 <400> 461 ggaagtgtag agtcaggaga tg 22 <210> 462 <211> 21 <212> DNA <213> CNAG_00670 Southern blot probe primer <400> 462 ggatgaaagg tgaaaggaca c 21 <210> 463 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 463 aacctttaaa tgggtagag 19 <210> 464 <211> 24 <212> DNA <213> STM common primer <400> 464 gcatgccctg cccctaagaa ttcg 24 <210> 465 <211> 18 <212> DNA <213> CNAG_05010 5 flanking region primer 1 <400> 465 tccttccttt ctaagccg 18 <210> 466 <211> 41 <212> DNA <213> CNAG_05010 5 flanking region primer 2 <400> 466 gctcactggc cgtcgtttta ctgtccccta cttttactcc g 41 <210> 467 <211> 41 <212> DNA <213> CNAG_05010 3 flanking region primer 1 <400> 467 catggtcata gctgtttcct gtgagggttt gataccttgt g 41 <210> 468 <211> 18 <212> DNA <213> CNAG_05010 3 flanking region primer 2 <400> 468 aagagcaaca tccgcaag 18 <210> 469 <211> 19 <212> DNA <213> CNAG_05010 diagnostic screening primer, pairing with B79 <400> 469 aacagccttg ccttacagc 19 <210> 470 <211> 19 <212> DNA <213> CNAG_05010 Southern blot probe primer <400> 470 agtggtgaag ggctaaatg 19 <210> 471 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 471 aacctttaaa tgggtagag 19 <210> 472 <211> 24 <212> DNA <213> STM common primer <400> 472 gcatgccctg cccctaagaa ttcg 24 <210> 473 <211> 18 <212> DNA <213> CNAG_04345 5 flanking region primer 1 <400> 473 aaaaggcagt cgtcttcg 18 <210> 474 <211> 40 <212> DNA <213> CNAG_04345 5 flanking region primer 2 <400> 474 ctggccgtcg ttttaccctt ttagatactc acacaagccc 40 <210> 475 <211> 36 <212> DNA <213> CNAG_04345 3 flanking region primer 1 <400> 475 gtcatagctg tttcctgttc tgggatgact tcacgc 36 <210> 476 <211> 19 <212> DNA <213> CNAG_04345 3 flanking region primer 2 <400> 476 ttactgctcc tggtttggg 19 <210> 477 <211> 18 <212> DNA <213> CNAG_04345 diagnostic screening primer, pairing with B79 <400> 477 cctgcttcac aaaatggc 18 <210> 478 <211> 22 <212> DNA <213> CNAG_04345 Southern blot probe primer <400> 478 gagtagtggt gtcagtgtga tg 22 <210> 479 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 479 ccatagaact agctaaagca 20 <210> 480 <211> 24 <212> DNA <213> STM common primer <400> 480 gcatgccctg cccctaagaa ttcg 24 <210> 481 <211> 19 <212> DNA <213> CNAG_02774 5 flanking region primer 1 <400> 481 ttgagccagc ctaagcaag 19 <210> 482 <211> 35 <212> DNA <213> CNAG_02774 5 flanking region primer 2 <400> 482 ctggccgtcg ttttactctt cttttctgcg gtcac 35 <210> 483 <211> 36 <212> DNA <213> CNAG_02774 3 flanking region primer 1 <400> 483 gtcatagctg tttcctggct tgatttcttg gaccac 36 <210> 484 <211> 19 <212> DNA <213> CNAG_02774 3 flanking region primer 2 <400> 484 cgagataaga agcaacgcc 19 <210> 485 <211> 19 <212> DNA <213> CNAG_02774 diagnostic screening primer, pairing with B79 <400> 485 tgccagccaa cctaatctg 19 <210> 486 <211> 20 <212> DNA <213> CNAG_02774 Southern blot probe primer <400> 486 gagaggcagg gagataactc 20 <210> 487 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 487 atgtaggtag ggtgataggt 20 <210> 488 <211> 24 <212> DNA <213> STM common primer <400> 488 gcatgccctg cccctaagaa ttcg 24 <210> 489 <211> 20 <212> DNA <213> CNAG_06425 5 flanking region primer 1 <400> 489 ctttcacgaa cgcttgattc 20 <210> 490 <211> 35 <212> DNA <213> CNAG_06425 5 flanking region primer 2 <400> 490 ctggccgtcg ttttacttgg acagtttttc ggcgg 35 <210> 491 <211> 36 <212> DNA <213> CNAG_06425 3 flanking region primer 1 <400> 491 gtcatagctg tttcctgatg tgcctgcgga tttctc 36 <210> 492 <211> 19 <212> DNA <213> CNAG_06425 3 flanking region primer 2 <400> 492 tgagaccgct gaaagttcc 19 <210> 493 <211> 19 <212> DNA <213> CNAG_06425 diagnostic screening primer, pairing with B79 <400> 493 taatcgccga caaagggtg 19 <210> 494 <211> 18 <212> DNA <213> CNAG_06425 Southern blot probe primer <400> 494 cccttttttt gtgcgacc 18 <210> 495 <211> 24 <212> DNA <213> NAT#288 STM primer <400> 495 ctatccaact agacctctag ctac 24 <210> 496 <211> 24 <212> DNA <213> STM common primer <400> 496 gcatgccctg cccctaagaa ttcg 24 <210> 497 <211> 22 <212> DNA <213> CNAG_05255 5 flanking region primer 1 <400> 497 atggtcaccg tcacaaggaa gc 22 <210> 498 <211> 40 <212> DNA <213> CNAG_05255 5 flanking region primer 2 <400> 498 ctggccgtcg ttttactaca cattcggctt ggttgcggac 40 <210> 499 <211> 38 <212> DNA <213> CNAG_05255 3 flanking region primer 1 <400> 499 gtcatagctg tttcctgcga ggtttgtatc ccagtcag 38 <210> 500 <211> 20 <212> DNA <213> CNAG_05255 3 flanking region primer 2 <400> 500 aggggctatc tctgcttcac 20 <210> 501 <211> 24 <212> DNA <213> CNAG_05255 diagnostic screening primer, pairing with B79 <400> 501 ttctgtgccc acctctactc tcag 24 <210> 502 <211> 20 <212> DNA <213> CNAG_05255 Southern blot probe primer <400> 502 accaaaactc gtcgtaggcg 20 <210> 503 <211> 24 <212> DNA <213> NAT#288 STM primer <400> 503 ctatccaact agacctctag ctac 24 <210> 504 <211> 24 <212> DNA <213> STM common primer <400> 504 gcatgccctg cccctaagaa ttcg 24 <210> 505 <211> 19 <212> DNA <213> CNAG_05049 5 flanking region primer 1 <400> 505 atcaggacga gtggcattg 19 <210> 506 <211> 39 <212> DNA <213> CNAG_05049 5 flanking region primer 2 <400> 506 tcactggccg tcgttttacg agtcttcttt gggcatagc 39 <210> 507 <211> 40 <212> DNA <213> CNAG_05049 3 flanking region primer 1 <400> 507 catggtcata gctgtttcct gagtggatga tgaggactcg 40 <210> 508 <211> 21 <212> DNA <213> CNAG_05049 3 flanking region primer 2 <400> 508 gtttcctatc ttcagtcgga g 21 <210> 509 <211> 20 <212> DNA <213> CNAG_05049 diagnostic screening primer, pairing with B79 <400> 509 gatagcagag gagaaaaggc 20 <210> 510 <211> 18 <212> DNA <213> CNAG_05049 Southern blot probe primer <400> 510 tatttggggt tgaggctg 18 <210> 511 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 511 ctatcgacca accaacacag 20 <210> 512 <211> 24 <212> DNA <213> STM common primer <400> 512 gcatgccctg cccctaagaa ttcg 24 <210> 513 <211> 18 <212> DNA <213> CNAG_05222 5 flanking region primer 1 <400> 513 gagggtcctt tggttttg 18 <210> 514 <211> 37 <212> DNA <213> CNAG_05222 5 flanking region primer 2 <400> 514 tcactggccg tcgttttact ttcacactca cggttgg 37 <210> 515 <211> 39 <212> DNA <213> CNAG_05222 3 flanking region primer 1 <400> 515 catggtcata gctgtttcct ggttgcccct tttgacaag 39 <210> 516 <211> 19 <212> DNA <213> CNAG_05222 3 flanking region primer 2 <400> 516 aacttacggc gacaacaac 19 <210> 517 <211> 18 <212> DNA <213> CNAG_05222 diagnostic screening primer, pairing with B79 <400> 517 tccacataat agccaccg 18 <210> 518 <211> 20 <212> DNA <213> CNAG_05222 Southern blot probe primer <400> 518 gctatgttca ggagcgatac 20 <210> 519 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 519 ctatcgacca accaacacag 20 <210> 520 <211> 24 <212> DNA <213> STM common primer <400> 520 gcatgccctg cccctaagaa ttcg 24 <210> 521 <211> 18 <212> DNA <213> CNAG_04130 5 flanking region primer 1 <400> 521 cattgacgcc atagcaag 18 <210> 522 <211> 38 <212> DNA <213> CNAG_04130 5 flanking region primer 2 <400> 522 tcactggccg tcgttttacg tgtttggaat gcggtttc 38 <210> 523 <211> 41 <212> DNA <213> CNAG_04130 3 flanking region primer 1 <400> 523 catggtcata gctgtttcct gtggatgagg ttgaggggaa c 41 <210> 524 <211> 21 <212> DNA <213> CNAG_04130 3 flanking region primer 2 <400> 524 gtagtgtaag ggcaggaaga g 21 <210> 525 <211> 21 <212> DNA <213> CNAG_04130 diagnostic screening primer, pairing with B79 <400> 525 ttctactcat aagggctacg c 21 <210> 526 <211> 21 <212> DNA <213> CNAG_04130 Southern blot probe primer <400> 526 ttctactcat aagggctacg c 21 <210> 527 <211> 19 <212> DNA <213> CNAG_00193 5 flanking region primer 1 <400> 527 gggcagaaat agttggcag 19 <210> 528 <211> 34 <212> DNA <213> CNAG_00193 5 flanking region primer 2 <400> 528 ctggccgtcg ttttactttt tccccccgat gtag 34 <210> 529 <211> 36 <212> DNA <213> CNAG_00193 3 flanking region primer 1 <400> 529 gtcatagctg tttcctgttc ccgcaagaac agtgtc 36 <210> 530 <211> 20 <212> DNA <213> CNAG_00193 3 flanking region primer 2 <400> 530 ttcaggatgt ccaaactcag 20 <210> 531 <211> 19 <212> DNA <213> CNAG_00193 diagnostic screening primer, pairing with B79 <400> 531 aaggggtaag aaatcgggc 19 <210> 532 <211> 19 <212> DNA <213> CNAG_00193 Southern blot probe primer <400> 532 tgctgagaga cagggaatc 19 <210> 533 <211> 20 <212> DNA <213> NAT#227 STM primer <400> 533 tcgtggttta gagggagcgc 20 <210> 534 <211> 24 <212> DNA <213> STM common primer <400> 534 gcatgccctg cccctaagaa ttcg 24 <210> 535 <211> 20 <212> DNA <213> CNAG_02700 5 flanking region primer 1 <400> 535 tgaaatcaga ctccagcctc 20 <210> 536 <211> 38 <212> DNA <213> CNAG_02700 5 flanking region primer 2 <400> 536 tcactggccg tcgttttacg cgaccacatt tacactcg 38 <210> 537 <211> 41 <212> DNA <213> CNAG_02700 3 flanking region primer 1 <400> 537 catggtcata gctgtttcct ggccacagaa caataactgg g 41 <210> 538 <211> 19 <212> DNA <213> CNAG_02700 3 flanking region primer 2 <400> 538 gcgtatgaat aagcgttgg 19 <210> 539 <211> 18 <212> DNA <213> CNAG_02700 diagnostic screening primer, pairing with B79 <400> 539 agcaaaacag aagcagcg 18 <210> 540 <211> 18 <212> DNA <213> CNAG_02700 Southern blot probe primer <400> 540 agaaagcagc atttgggg 18 <210> 541 <211> 20 <212> DNA <213> NAT#230 STM primer <400> 541 atgtaggtag ggtgataggt 20 <210> 542 <211> 24 <212> DNA <213> STM common primer <400> 542 gcatgccctg cccctaagaa ttcg 24 <210> 543 <211> 19 <212> DNA <213> CNAG_04586 5 flanking region primer 1 <400> 543 tccatctccc aatccacag 19 <210> 544 <211> 38 <212> DNA <213> CNAG_04586 5 flanking region primer 2 <400> 544 tcactggccg tcgttttacg cttgattagg cggataag 38 <210> 545 <211> 41 <212> DNA <213> CNAG_04586 3 flanking region primer 1 <400> 545 catggtcata gctgtttcct gtggacgaaa agacccctac c 41 <210> 546 <211> 18 <212> DNA <213> CNAG_04586 3 flanking region primer 2 <400> 546 aacactggaa tgtgcgtg 18 <210> 547 <211> 20 <212> DNA <213> CNAG_04586 diagnostic screening primer, pairing with B79 <400> 547 ttggtggtgc ggttttgtgc 20 <210> 548 <211> 18 <212> DNA <213> CNAG_04586 Southern blot probe primer <400> 548 tgatgttggt tcgtgacc 18 <210> 549 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 549 acgcaccaga cacacaacca g 21 <210> 550 <211> 24 <212> DNA <213> STM common primer <400> 550 gcatgccctg cccctaagaa ttcg 24 <210> 551 <211> 18 <212> DNA <213> CNAG_05153 5 flanking region primer 1 <400> 551 aaaaccaagt ggaacccg 18 <210> 552 <211> 35 <212> DNA <213> CNAG_05153 5 flanking region primer 2 <400> 552 ctggccgtcg ttttactggt tgacgctgtt agcac 35 <210> 553 <211> 37 <212> DNA <213> CNAG_05153 3 flanking region primer 1 <400> 553 gtcatagctg tttcctgata ggcaggctgg tggaaac 37 <210> 554 <211> 19 <212> DNA <213> CNAG_05153 3 flanking region primer 2 <400> 554 accacatttg cttccgagc 19 <210> 555 <211> 21 <212> DNA <213> CNAG_05153 diagnostic screening primer, pairing with JOHE12579 <400> 555 tggctgaact gtaggttagt g 21 <210> 556 <211> 18 <212> DNA <213> CNAG_05153 Southern blot probe primer <400> 556 cggcgataag gtttttgc 18 <210> 557 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 557 accgacagct cgaacaagca agag 24 <210> 558 <211> 24 <212> DNA <213> STM common primer <400> 558 gcatgccctg cccctaagaa ttcg 24 <210> 559 <211> 20 <212> DNA <213> CNAG_02723 5 flanking region primer 1 <400> 559 gcttccttat cggtagggac 20 <210> 560 <211> 39 <212> DNA <213> CNAG_02723 5 flanking region primer 2 <400> 560 tcactggccg tcgttttacg actggacatt gacgaagac 39 <210> 561 <211> 40 <212> DNA <213> CNAG_02723 3 flanking region primer 1 <400> 561 catggtcata gctgtttcct gtggtgacag tgacgcttac 40 <210> 562 <211> 19 <212> DNA <213> CNAG_02723 3 flanking region primer 2 <400> 562 attgtcagga tgtgggttc 19 <210> 563 <211> 21 <212> DNA <213> CNAG_02723 diagnostic screening primer, pairing with B79 <400> 563 gcttactccc aaactcacac c 21 <210> 564 <211> 18 <212> DNA <213> CNAG_02723 Southern blot probe primer <400> 564 atggatttgg cagaacgg 18 <210> 565 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 565 caccctctat ctcgagaaag ctcc 24 <210> 566 <211> 24 <212> DNA <213> STM common primer <400> 566 gcatgccctg cccctaagaa ttcg 24 <210> 567 <211> 19 <212> DNA <213> CNAG_03741 5 flanking region primer 1 <400> 567 gccgtctcct gatgtaatc 19 <210> 568 <211> 36 <212> DNA <213> CNAG_03741 5 flanking region primer 2 <400> 568 ctggccgtcg ttttaccgac tgtcatcaac gactgg 36 <210> 569 <211> 36 <212> DNA <213> CNAG_03741 3 flanking region primer 1 <400> 569 ctggccgtcg ttttacgcgt caacaggaat aaaacc 36 <210> 570 <211> 22 <212> DNA <213> CNAG_03741 3 flanking region primer 2 <400> 570 ctgtgccttt ctccaagata ag 22 <210> 571 <211> 22 <212> DNA <213> CNAG_03741 diagnostic screening primer, pairing with B79 <400> 571 ggataagcag aacatagttg cg 22 <210> 572 <211> 21 <212> DNA <213> CNAG_03741 Southern blot probe primer <400> 572 tctcttcttc ttctgacctg c 21 <210> 573 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 573 gagatctttc gggaggtctg gatt 24 <210> 574 <211> 24 <212> DNA <213> STM common primer <400> 574 gcatgccctg cccctaagaa ttcg 24 <210> 575 <211> 18 <212> DNA <213> CNAG_07411 5 flanking region primer 1 <400> 575 agacccgcca aaacaatc 18 <210> 576 <211> 35 <212> DNA <213> CNAG_07411 5 flanking region primer 2 <400> 576 ctggccgtcg ttttacttgg acctacgctg acttc 35 <210> 577 <211> 37 <212> DNA <213> CNAG_07411 3 flanking region primer 1 <400> 577 gtcatagctg tttcctggac ttgcgttctg attttgg 37 <210> 578 <211> 20 <212> DNA <213> CNAG_07411 3 flanking region primer 2 <400> 578 gctgttatca agagaatgcc 20 <210> 579 <211> 19 <212> DNA <213> CNAG_07411 diagnostic screening primer, pairing with B79 <400> 579 acgactggct atcaagcag 19 <210> 580 <211> 19 <212> DNA <213> CNAG_07411 Southern blot probe primer <400> 580 tacgaggatt gagtctggc 19 <210> 581 <211> 24 <212> DNA <213> NAT#288 STM primer <400> 581 ctatccaact agacctctag ctac 24 <210> 582 <211> 24 <212> DNA <213> STM common primer <400> 582 gcatgccctg cccctaagaa ttcg 24 <210> 583 <211> 20 <212> DNA <213> CNAG_03849 5 flanking region primer 1 <400> 583 tagtggttcc cgtttgtgcg 20 <210> 584 <211> 40 <212> DNA <213> CNAG_03849 5 flanking region primer 2 <400> 584 gctcactggc cgtcgtttta ccttacgcct ttttccatcg 40 <210> 585 <211> 41 <212> DNA <213> CNAG_03849 3 flanking region primer 1 <400> 585 catggtcata gctgtttcct gtcaaacggc gattggtggt c 41 <210> 586 <211> 20 <212> DNA <213> CNAG_03849 3 flanking region primer 2 <400> 586 tggtgaaggg gaagattggg 20 <210> 587 <211> 19 <212> DNA <213> CNAG_03849 diagnostic screening primer, pairing with B79 <400> 587 aacaagccga ggaatgagg 19 <210> 588 <211> 19 <212> DNA <213> CNAG_03849 Southern blot probe primer <400> 588 tgagtgaacg aagagtgcc 19 <210> 589 <211> 20 <212> DNA <213> NAT#6 STM primer <400> 589 atagctacca cacgatagct 20 <210> 590 <211> 24 <212> DNA <213> STM common primer <400> 590 gcatgccctg cccctaagaa ttcg 24 <210> 591 <211> 19 <212> DNA <213> CNAG_04916 5 flanking region primer 1 <400> 591 ttgcgttgag acatcactg 19 <210> 592 <211> 43 <212> DNA <213> CNAG_04916 5 flanking region primer 2 <400> 592 gctcactggc cgtcgtttta cgctcctatg ttaccaatgt tgc 43 <210> 593 <211> 40 <212> DNA <213> CNAG_04916 3 flanking region primer 1 <400> 593 catggtcata gctgtttcct gacccatttg ctttcctggc 40 <210> 594 <211> 20 <212> DNA <213> CNAG_04916 3 flanking region primer 2 <400> 594 tctcaacgag tcgctttcgc 20 <210> 595 <211> 20 <212> DNA <213> CNAG_04916 diagnostic screening primer, pairing with B79 <400> 595 aaaagcatcc tgcccagcac 20 <210> 596 <211> 20 <212> DNA <213> CNAG_04916 Southern blot probe primer <400> 596 gttgaaagta gttgcgttgg 20 <210> 597 <211> 20 <212> DNA <213> NAT#212 STM primer <400> 597 agagcgatcg cgttatagat 20 <210> 598 <211> 24 <212> DNA <213> STM common primer <400> 598 gcatgccctg cccctaagaa ttcg 24 <210> 599 <211> 19 <212> DNA <213> CNAG_04012 5 flanking region primer 1 <400> 599 ttggtggagt gattgaccc 19 <210> 600 <211> 36 <212> DNA <213> CNAG_04012 5 flanking region primer 2 <400> 600 ctggccgtcg ttttacaatc aaaccagaag gagagc 36 <210> 601 <211> 39 <212> DNA <213> CNAG_04012 3 flanking region primer 1 <400> 601 gtcatagctg tttcctgagg aggataaagg aagagagtg 39 <210> 602 <211> 19 <212> DNA <213> CNAG_04012 3 flanking region primer 2 <400> 602 ttatgctggt ggtgcctac 19 <210> 603 <211> 19 <212> DNA <213> CNAG_04012 diagnostic screening primer, pairing with B79 <400> 603 accgaagagg atgaggttg 19 <210> 604 <211> 20 <212> DNA <213> CNAG_04012 Southern blot probe primer <400> 604 cagtggtggt tgagattgac 20 <210> 605 <211> 20 <212> DNA <213> NAT#212 STM primer <400> 605 agagcgatcg cgttatagat 20 <210> 606 <211> 24 <212> DNA <213> STM common primer <400> 606 gcatgccctg cccctaagaa ttcg 24 <210> 607 <211> 20 <212> DNA <213> CNAG_04090 5 flanking region primer 1 <400> 607 gagtagagga gtggattggg 20 <210> 608 <211> 39 <212> DNA <213> CNAG_04090 5 flanking region primer 2 <400> 608 gctcactggc cgtcgtttta cccattgttc aagcagagc 39 <210> 609 <211> 42 <212> DNA <213> CNAG_04090 3 flanking region primer 1 <400> 609 catggtcata gctgtttcct gggcttactg ctatcttgat gc 42 <210> 610 <211> 24 <212> DNA <213> CNAG_04090 3 flanking region primer 2 <400> 610 gaaagaggaa ctacatactg ggtc 24 <210> 611 <211> 20 <212> DNA <213> CNAG_04090 diagnostic screening primer, pairing with B79 <400> 611 gtcttttgct ccttgaaacc 20 <210> 612 <211> 20 <212> DNA <213> CNAG_04090 Southern blot probe primer <400> 612 accaggtgta aacccagtcc 20 <210> 613 <211> 20 <212> DNA <213> NAT#220 STM primer <400> 613 cagatctcga acgataccca 20 <210> 614 <211> 24 <212> DNA <213> STM common primer <400> 614 gcatgccctg cccctaagaa ttcg 24 <210> 615 <211> 21 <212> DNA <213> CNAG_05375 5 flanking region primer 1 <400> 615 ggaagcatta ctgtcccata g 21 <210> 616 <211> 43 <212> DNA <213> CNAG_05375 5 flanking region primer 2 <400> 616 gctcactggc cgtcgtttta cagaaacagg agagagagta gcc 43 <210> 617 <211> 39 <212> DNA <213> CNAG_05375 3 flanking region primer 1 <400> 617 catggtcata gctgtttcct gtcttggatt tcaacgggg 39 <210> 618 <211> 20 <212> DNA <213> CNAG_05375 3 flanking region primer 2 <400> 618 gtagttccgt tatttgctgg 20 <210> 619 <211> 22 <212> DNA <213> CNAG_05375 diagnostic screening primer, pairing with B79 <400> 619 ggttgaagat ggacgagtag ac 22 <210> 620 <211> 20 <212> DNA <213> CNAG_05375 Southern blot probe primer <400> 620 cattcacctc ccttagaagc 20 <210> 621 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 621 aacctttaaa tgggtagag 19 <210> 622 <211> 24 <212> DNA <213> STM common primer <400> 622 gcatgccctg cccctaagaa ttcg 24 <210> 623 <211> 20 <212> DNA <213> CNAG_07797 5 flanking region primer 1 <400> 623 agggagaaca ccacaggatg 20 <210> 624 <211> 38 <212> DNA <213> CNAG_07797 5 flanking region primer 2 <400> 624 tcactggccg tcgttttaca ggaatggagg tggtgaac 38 <210> 625 <211> 40 <212> DNA <213> CNAG_07797 3 flanking region primer 1 <400> 625 catggtcata gctgtttcct gggaaagtca gtgatgctcc 40 <210> 626 <211> 18 <212> DNA <213> CNAG_07797 3 flanking region primer 2 <400> 626 gctttcttcc ttttcccg 18 <210> 627 <211> 18 <212> DNA <213> CNAG_07797 diagnostic screening primer, pairing with B79 <400> 627 atcaatggtt tcgtcggc 18 <210> 628 <211> 18 <212> DNA <213> CNAG_07797 Southern blot probe primer <400> 628 ggggcgttga aaatgatg 18 <210> 629 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 629 gagagatccc aacatcacgc 20 <210> 630 <211> 24 <212> DNA <213> STM common primer <400> 630 gcatgccctg cccctaagaa ttcg 24 <210> 631 <211> 22 <212> DNA <213> CNAG_05538 5 flanking region primer 1 <400> 631 ggtctcaaca aagcacttac tg 22 <210> 632 <211> 40 <212> DNA <213> CNAG_05538 5 flanking region primer 2 <400> 632 tcactggccg tcgttttacg taatagtcag gcggtttctc 40 <210> 633 <211> 41 <212> DNA <213> CNAG_05538 3 flanking region primer 1 <400> 633 catggtcata gctgtttcct gagttaccag aggaggagtg g 41 <210> 634 <211> 20 <212> DNA <213> CNAG_05538 3 flanking region primer 2 <400> 634 gattctcttt accctcagcc 20 <210> 635 <211> 20 <212> DNA <213> CNAG_05538 diagnostic screening primer, pairing with B79 <400> 635 tccgattcta ttgactctgc 20 <210> 636 <211> 20 <212> DNA <213> CNAG_05538 Southern blot probe primer <400> 636 gtatcatcca ttttcctggc 20 <210> 637 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 637 ggtgttggat cggggtggat 20 <210> 638 <211> 24 <212> DNA <213> STM common primer <400> 638 gcatgccctg cccctaagaa ttcg 24 <210> 639 <211> 18 <212> DNA <213> CNAG_00332 5 flanking region primer 1 <400> 639 tgattgtcgt ccttgtgg 18 <210> 640 <211> 39 <212> DNA <213> CNAG_00332 5 flanking region primer 2 <400> 640 gctcactggc cgtcgtttta ctttgcgtga ctgttctgc 39 <210> 641 <211> 42 <212> DNA <213> CNAG_00332 3 flanking region primer 1 <400> 641 catggtcata gctgtttcct gcggtttgcg aagatacaga ac 42 <210> 642 <211> 18 <212> DNA <213> CNAG_00332 3 flanking region primer 2 <400> 642 tatggatgcg tgggaatg 18 <210> 643 <211> 18 <212> DNA <213> CNAG_00332 diagnostic screening primer, pairing with B79 <400> 643 gccagaccaa tcaaatgc 18 <210> 644 <211> 19 <212> DNA <213> CNAG_00332 Southern blot probe primer <400> 644 gaagaagggt tgaatgtcg 19 <210> 645 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 645 accgacagct cgaacaagca agag 24 <210> 646 <211> 24 <212> DNA <213> STM common primer <400> 646 gcatgccctg cccctaagaa ttcg 24 <210> 647 <211> 19 <212> DNA <213> CNAG_04023 5 flanking region primer 1 <400> 647 agcaggatag acagcgaag 19 <210> 648 <211> 35 <212> DNA <213> CNAG_04023 5 flanking region primer 2 <400> 648 ctggccgtcg ttttactgtc cttgataggt cggag 35 <210> 649 <211> 35 <212> DNA <213> CNAG_04023 3 flanking region primer 1 <400> 649 gtcatagctg tttcctgtgg aacacattgg aggag 35 <210> 650 <211> 18 <212> DNA <213> CNAG_04023 3 flanking region primer 2 <400> 650 aatgggtgta tgatggcg 18 <210> 651 <211> 19 <212> DNA <213> CNAG_04023 diagnostic screening primer, pairing with B79 <400> 651 cagaacgcat tgtttgtgc 19 <210> 652 <211> 19 <212> DNA <213> CNAG_04023 Southern blot probe primer <400> 652 gtagtaaacg ccttgagcc 19 <210> 653 <211> 20 <212> DNA <213> CNAG_05311 5 flanking region primer 1 <400> 653 caacaagttc catcacatcg 20 <210> 654 <211> 40 <212> DNA <213> CNAG_05311 5 flanking region primer 2 <400> 654 gctcactggc cgtcgtttta cgggtaaaac ctggcttagg 40 <210> 655 <211> 39 <212> DNA <213> CNAG_05311 3 flanking region primer 1 <400> 655 catggtcata gctgtttcct gggatttgga tgaacctgg 39 <210> 656 <211> 21 <212> DNA <213> CNAG_05311 3 flanking region primer 2 <400> 656 gagaaagagg tttgtcgtga g 21 <210> 657 <211> 18 <212> DNA <213> CNAG_05311 diagnostic screening primer, pairing with B79 <400> 657 ttctactttc cgccaagg 18 <210> 658 <211> 19 <212> DNA <213> CNAG_05311 Southern blot probe primer <400> 658 gagatgggtc actaatcgc 19 <210> 659 <211> 19 <212> DNA <213> CNAG_03086 5 flanking region primer 1 <400> 659 aatctccttt gcccgactc 19 <210> 660 <211> 38 <212> DNA <213> CNAG_03086 5 flanking region primer 2 <400> 660 tcactggccg tcgttttaca taccttgtct ttgcgggc 38 <210> 661 <211> 41 <212> DNA <213> CNAG_03086 3 flanking region primer 1 <400> 661 catggtcata gctgtttcct gattgaaggt ggcagtagat g 41 <210> 662 <211> 18 <212> DNA <213> CNAG_03086 3 flanking region primer 2 <400> 662 aaaatgtggg cggatgag 18 <210> 663 <211> 19 <212> DNA <213> CNAG_03086 diagnostic screening primer, pairing with B79 <400> 663 tcagtcggga agagtttgc 19 <210> 664 <211> 19 <212> DNA <213> CNAG_03086 Southern blot probe primer <400> 664 aggcgagagt aagaatgcg 19 <210> 665 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 665 atatggatgt ttttagcgag 20 <210> 666 <211> 24 <212> DNA <213> STM common primer <400> 666 gcatgccctg cccctaagaa ttcg 24 <210> 667 <211> 18 <212> DNA <213> CNAG_03336 5 flanking region primer 1 <400> 667 gtgccttatt ccaatggg 18 <210> 668 <211> 34 <212> DNA <213> CNAG_03336 5 flanking region primer 2 <400> 668 ctggccgtcg ttttacttct tgctgcctgt ttgg 34 <210> 669 <211> 35 <212> DNA <213> CNAG_03336 3 flanking region primer 1 <400> 669 ctggccgtcg ttttacccaa caataatcac gcagg 35 <210> 670 <211> 20 <212> DNA <213> CNAG_03336 3 flanking region primer 2 <400> 670 ccatcctttt tttagcggtc 20 <210> 671 <211> 19 <212> DNA <213> CNAG_03336 diagnostic screening primer, pairing with B79 <400> 671 cgaggtgatt gatgtgagc 19 <210> 672 <211> 20 <212> DNA <213> CNAG_03336 Southern blot probe primer <400> 672 ttcaagagcg gactggacag 20 <210> 673 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 673 gatctctcgc gcttggggga 20 <210> 674 <211> 24 <212> DNA <213> STM common primer <400> 674 gcatgccctg cccctaagaa ttcg 24 <210> 675 <211> 19 <212> DNA <213> CNAG_03229 5 flanking region primer 1 <400> 675 gtgggttact ttgggtgag 19 <210> 676 <211> 37 <212> DNA <213> CNAG_03229 5 flanking region primer 2 <400> 676 tcactggccg tcgttttaca aaagggactt gctgtcg 37 <210> 677 <211> 40 <212> DNA <213> CNAG_03229 3 flanking region primer 1 <400> 677 catggtcata gctgtttcct ggaaagaaag gaaatcgccg 40 <210> 678 <211> 18 <212> DNA <213> CNAG_03229 3 flanking region primer 2 <400> 678 tggaagatgg tgacgaag 18 <210> 679 <211> 18 <212> DNA <213> CNAG_03229 diagnostic screening primer, pairing with B79 <400> 679 ttgcgatttc cgactacc 18 <210> 680 <211> 18 <212> DNA <213> CNAG_03229 Southern blot probe primer <400> 680 tggaggcgaa taatggtc 18 <210> 681 <211> 20 <212> DNA <213> NAT#227 STM primer <400> 681 tcgtggttta gagggagcgc 20 <210> 682 <211> 24 <212> DNA <213> STM common primer <400> 682 gcatgccctg cccctaagaa ttcg 24 <210> 683 <211> 19 <212> DNA <213> CNAG_01883 5 flanking region primer 1 <400> 683 tactcgctgc ttagggttc 19 <210> 684 <211> 35 <212> DNA <213> CNAG_01883 5 flanking region primer 2 <400> 684 ctggccgtcg ttttacatgg catactccag gtctg 35 <210> 685 <211> 36 <212> DNA <213> CNAG_01883 3 flanking region primer 1 <400> 685 gtcatagctg tttcctgccc tccatttact tctccg 36 <210> 686 <211> 22 <212> DNA <213> CNAG_01883 3 flanking region primer 2 <400> 686 tagcagtagg gctcctgatt cg 22 <210> 687 <211> 19 <212> DNA <213> CNAG_01883 diagnostic screening primer, pairing with B79 <400> 687 ttggaggtct ttatcagcg 19 <210> 688 <211> 19 <212> DNA <213> CNAG_01883 Southern blot probe primer <400> 688 ggagaagtaa atggagggg 19 <210> 689 <211> 24 <212> DNA <213> NAT#125 STM primer <400> 689 cgctacagcc agcgcgcgca agcg 24 <210> 690 <211> 24 <212> DNA <213> STM common primer <400> 690 gcatgccctg cccctaagaa ttcg 24 <210> 691 <211> 20 <212> DNA <213> CNAG_04837 5 flanking region primer 1 <400> 691 tgaaacagtg aatcaggcac 20 <210> 692 <211> 40 <212> DNA <213> CNAG_04837 5 flanking region primer 2 <400> 692 gctcactggc cgtcgtttta ctcagtgtca acggtcattg 40 <210> 693 <211> 42 <212> DNA <213> CNAG_04837 3 flanking region primer 1 <400> 693 catggtcata gctgtttcct ggaaaggggt agggataagt tg 42 <210> 694 <211> 20 <212> DNA <213> CNAG_04837 3 flanking region primer 2 <400> 694 actgaacgac aagagacgac 20 <210> 695 <211> 18 <212> DNA <213> CNAG_04837 diagnostic screening primer, pairing with B79 <400> 695 taagagaaaa ggggctgc 18 <210> 696 <211> 21 <212> DNA <213> CNAG_04837 Southern blot probe primer 1 <400> 696 tgtccccatc aactctacaa c 21 <210> 697 <211> 20 <212> DNA <213> CNAG_04837 Southern blot probe primer 2 <400> 697 cttctgcttt caccattagc 20 <210> 698 <211> 21 <212> DNA <213> NAT#146 STM primer <400> 698 actagccccc cctcaccacc t 21 <210> 699 <211> 24 <212> DNA <213> STM common primer <400> 699 gcatgccctg cccctaagaa ttcg 24 <210> 700 <211> 19 <212> DNA <213> CNAG_00514 5 flanking region primer 1 <400> 700 ggtgtttcca tacgaaccg 19 <210> 701 <211> 38 <212> DNA <213> CNAG_00514 5 flanking region primer 2 <400> 701 ctggccgtcg ttttacggag ccatttctta cttatgcg 38 <210> 702 <211> 35 <212> DNA <213> CNAG_00514 3 flanking region primer 1 <400> 702 gtcatagctg tttcctgaat cccaaggacc atctg 35 <210> 703 <211> 20 <212> DNA <213> CNAG_00514 3 flanking region primer 2 <400> 703 gctgagaagg agaggatagc 20 <210> 704 <211> 20 <212> DNA <213> CNAG_00514 diagnostic screening primer, pairing with B79 <400> 704 gtggttgaaa gtattaggcg 20 <210> 705 <211> 19 <212> DNA <213> CNAG_00514 Southern blot probe primer <400> 705 gaggattggt gatggactg 19 <210> 706 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 706 caccctctat ctcgagaaag ctcc 24 <210> 707 <211> 24 <212> DNA <213> STM common primer <400> 707 gcatgccctg cccctaagaa ttcg 24 <210> 708 <211> 20 <212> DNA <213> CNAG_03409 5 flanking region primer 1 <400> 708 cctcaccgat tcattcactg 20 <210> 709 <211> 38 <212> DNA <213> CNAG_03409 5 flanking region primer 2 <400> 709 ctggccgtcg ttttactttt ctcccttctt cgcccgac 38 <210> 710 <211> 37 <212> DNA <213> CNAG_03409 3 flanking region primer 1 <400> 710 gtcatagctg tttcctgagg ttcaagccca gactcag 37 <210> 711 <211> 20 <212> DNA <213> CNAG_03409 3 flanking region primer 2 <400> 711 agtatctacg gtgctggtgc 20 <210> 712 <211> 20 <212> DNA <213> CNAG_03409 diagnostic screening primer, pairing with B79 <400> 712 tgaggatgat gactggggac 20 <210> 713 <211> 20 <212> DNA <213> CNAG_03409 Southern blot probe primer 1 <400> 713 acttccaatg ctcaaccccg 20 <210> 714 <211> 20 <212> DNA <213> CNAG_03409 Southern blot probe primer 2 <400> 714 gcaggacaag aaggagagtg 20 <210> 715 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 715 caccctctat ctcgagaaag ctcc 24 <210> 716 <211> 24 <212> DNA <213> STM common primer <400> 716 gcatgccctg cccctaagaa ttcg 24 <210> 717 <211> 19 <212> DNA <213> CNAG_05392 5 flanking region primer 1 <400> 717 aagttgtgaa accaccagg 19 <210> 718 <211> 38 <212> DNA <213> CNAG_05392 5 flanking region primer 2 <400> 718 tcactggccg tcgttttact caccctgtag atgtgtgg 38 <210> 719 <211> 42 <212> DNA <213> CNAG_05392 3 flanking region primer 1 <400> 719 catggtcata gctgtttcct gggagtccct gtgaagtaaa ag 42 <210> 720 <211> 20 <212> DNA <213> CNAG_05392 3 flanking region primer 2 <400> 720 tgagccaaag tagcagtcag 20 <210> 721 <211> 19 <212> DNA <213> CNAG_05392 diagnostic screening primer, pairing with B79 <400> 721 gaatggacct tggaaaacg 19 <210> 722 <211> 20 <212> DNA <213> CNAG_05392 Southern blot probe primer <400> 722 cggtctgtgt gcttcttcac 20 <210> 723 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 723 gatctctcgc gcttggggga 20 <210> 724 <211> 24 <212> DNA <213> STM common primer <400> 724 gcatgccctg cccctaagaa ttcg 24 <210> 725 <211> 20 <212> DNA <213> CNAG_04895 5 flanking region primer 1 <400> 725 ttcgtagtta gccagcagag 20 <210> 726 <211> 38 <212> DNA <213> CNAG_04895 5 flanking region primer 2 <400> 726 tcactggccg tcgttttacg ttcttgatgt ggaggagc 38 <210> 727 <211> 40 <212> DNA <213> CNAG_04895 3 flanking region primer 1 <400> 727 catggtcata gctgtttcct gcccaaccct tatgcgtatg 40 <210> 728 <211> 18 <212> DNA <213> CNAG_04895 3 flanking region primer 2 <400> 728 agcgtctccc tcaaaaac 18 <210> 729 <211> 18 <212> DNA <213> CNAG_04895 diagnostic screening primer, pairing with B79 <400> 729 ttagcgaccg aaaaaggg 18 <210> 730 <211> 19 <212> DNA <213> CNAG_04895 Southern blot probe primer <400> 730 aagatgacgc agaactggg 19 <210> 731 <211> 20 <212> DNA <213> NAT#204 STM primer <400> 731 gatctctcgc gcttggggga 20 <210> 732 <211> 24 <212> DNA <213> STM common primer <400> 732 gcatgccctg cccctaagaa ttcg 24 <210> 733 <211> 19 <212> DNA <213> CNAG_04176 5 flanking region primer 1 <400> 733 ttgacacggg gactaatcg 19 <210> 734 <211> 40 <212> DNA <213> CNAG_04176 5 flanking region primer 2 <400> 734 gctcactggc cgtcgtttta cgattagccc agtcttagcg 40 <210> 735 <211> 41 <212> DNA <213> CNAG_04176 3 flanking region primer 1 <400> 735 catggtcata gctgtttcct gagaagcagg gagcaaaacg g 41 <210> 736 <211> 19 <212> DNA <213> CNAG_04176 3 flanking region primer 2 <400> 736 ccgaaaagtt gagcaagac 19 <210> 737 <211> 23 <212> DNA <213> CNAG_04176 diagnostic screening primer, pairing with B79 <400> 737 cgatagcctt tgtagtaggt ctc 23 <210> 738 <211> 20 <212> DNA <213> CNAG_04176 Southern blot probe primer <400> 738 gggatttgat tatctggagg 20 <210> 739 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 739 tatccccctc tccgctctct agca 24 <210> 740 <211> 24 <212> DNA <213> STM common primer <400> 740 gcatgccctg cccctaagaa ttcg 24 <210> 741 <211> 19 <212> DNA <213> CNAG_07724 5 flanking region primer 1 <400> 741 caaagaaaac gctcactcc 19 <210> 742 <211> 40 <212> DNA <213> CNAG_07724 5 flanking region primer 2 <400> 742 tcactggccg tcgttttacg gatagaacaa gacaagagcg 40 <210> 743 <211> 41 <212> DNA <213> CNAG_07724 3 flanking region primer 1 <400> 743 catggtcata gctgtttcct gacttcagtc gttaccagca c 41 <210> 744 <211> 19 <212> DNA <213> CNAG_07724 3 flanking region primer 2 <400> 744 tctttgagca catcaggtg 19 <210> 745 <211> 19 <212> DNA <213> CNAG_07724 diagnostic screening primer, pairing with B79 <400> 745 ccgtcattat cgtgtctcc 19 <210> 746 <211> 19 <212> DNA <213> CNAG_07724 Southern blot probe primer <400> 746 gcttcggagg agttgatag 19 <210> 747 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 747 tatccccctc tccgctctct agca 24 <210> 748 <211> 24 <212> DNA <213> STM common primer <400> 748 gcatgccctg cccctaagaa ttcg 24 <210> 749 <211> 19 <212> DNA <213> CNAG_06751 5 flanking region primer 1 <400> 749 aaaaacggct gaaggctcg 19 <210> 750 <211> 40 <212> DNA <213> CNAG_06751 5 flanking region primer 2 <400> 750 gctcactggc cgtcgtttta ctggcgagtt ggtatctgag 40 <210> 751 <211> 40 <212> DNA <213> CNAG_06751 3 flanking region primer 1 <400> 751 catggtcata gctgtttcct gaagatgaga gagaaggggg 40 <210> 752 <211> 20 <212> DNA <213> CNAG_06751 3 flanking region primer 2 <400> 752 agaagtggta tgaccgtttg 20 <210> 753 <211> 19 <212> DNA <213> CNAG_06751 diagnostic screening primer, pairing with B79 <400> 753 tccatacatt gttgagggc 19 <210> 754 <211> 18 <212> DNA <213> CNAG_06751 Southern blot probe primer <400> 754 attgcccaag cggataag 18 <210> 755 <211> 22 <212> DNA <213> NAT#208 STM primer <400> 755 tggtcgcggg agatcgtggt tt 22 <210> 756 <211> 24 <212> DNA <213> STM common primer <400> 756 gcatgccctg cccctaagaa ttcg 24 <210> 757 <211> 18 <212> DNA <213> CNAG_01438 5 flanking region primer 1 <400> 757 atcacacgca atcccatc 18 <210> 758 <211> 39 <212> DNA <213> CNAG_01438 5 flanking region primer 2 <400> 758 gctcactggc cgtcgtttta cagcaagatg gaaggaagc 39 <210> 759 <211> 39 <212> DNA <213> CNAG_01438 3 flanking region primer 1 <400> 759 catggtcata gctgtttcct ggcgacgaaa tcataaggc 39 <210> 760 <211> 19 <212> DNA <213> CNAG_01438 3 flanking region primer 2 <400> 760 tgagaactgg acgaactgc 19 <210> 761 <211> 18 <212> DNA <213> CNAG_01438 diagnostic screening primer, pairing with B79 <400> 761 aatgatgcgt cggacttc 18 <210> 762 <211> 20 <212> DNA <213> CNAG_01438 Southern blot probe primer <400> 762 ggaagagaaa gacttgcgag 20 <210> 763 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 763 acagctccaa acctcgctaa acag 24 <210> 764 <211> 24 <212> DNA <213> STM common primer <400> 764 gcatgccctg cccctaagaa ttcg 24 <210> 765 <211> 20 <212> DNA <213> CNAG_00791 5 flanking region primer 1 <400> 765 gcatccattt tatcggttcg 20 <210> 766 <211> 41 <212> DNA <213> CNAG_00791 5 flanking region primer 2 <400> 766 gctcactggc cgtcgtttta cttctgcttg aaggagtcgg g 41 <210> 767 <211> 40 <212> DNA <213> CNAG_00791 3 flanking region primer 1 <400> 767 catggtcata gctgtttcct gcggttatct tgcgtaaagc 40 <210> 768 <211> 19 <212> DNA <213> CNAG_00791 3 flanking region primer 2 <400> 768 tgtctcaccc atcacgaac 19 <210> 769 <211> 18 <212> DNA <213> CNAG_00791 diagnostic screening primer, pairing with B79 <400> 769 attcgctcag gcagtttc 18 <210> 770 <211> 19 <212> DNA <213> CNAG_00791 Southern blot probe primer <400> 770 gcgatgatga ggactcttg 19 <210> 771 <211> 21 <212> DNA <213> NAT#146 STM primer <400> 771 actagccccc cctcaccacc t 21 <210> 772 <211> 24 <212> DNA <213> STM common primer <400> 772 gcatgccctg cccctaagaa ttcg 24 <210> 773 <211> 19 <212> DNA <213> CNAG_02241 5 flanking region primer 1 <400> 773 atagcaaacc catacctcg 19 <210> 774 <211> 37 <212> DNA <213> CNAG_02241 5 flanking region primer 2 <400> 774 tcactggccg tcgttttact ttgtttggta gtggggg 37 <210> 775 <211> 40 <212> DNA <213> CNAG_02241 3 flanking region primer 1 <400> 775 catggtcata gctgtttcct gcgagaactc tggagcattc 40 <210> 776 <211> 21 <212> DNA <213> CNAG_02241 3 flanking region primer 2 <400> 776 gatgtgttgt ggactgtatg c 21 <210> 777 <211> 20 <212> DNA <213> CNAG_02241 diagnostic screening primer, pairing with B79 <400> 777 tcccatcata ggagtggacg 20 <210> 778 <211> 20 <212> DNA <213> CNAG_02241 Southern blot probe primer <400> 778 cttgggagca aataagtgtg 20 <210> 779 <211> 20 <212> DNA <213> NAT#219 STM primer <400> 779 ccctaaaacc ctacagcaat 20 <210> 780 <211> 24 <212> DNA <213> STM common primer <400> 780 gcatgccctg cccctaagaa ttcg 24 <210> 781 <211> 19 <212> DNA <213> CNAG_02566 5 flanking region primer 1 <400> 781 tcgcacggca taggattag 19 <210> 782 <211> 38 <212> DNA <213> CNAG_02566 5 flanking region primer 2 <400> 782 tcactggccg tcgttttaca tgccgccaga atgttgag 38 <210> 783 <211> 40 <212> DNA <213> CNAG_02566 3 flanking region primer 1 <400> 783 catggtcata gctgtttcct gtgatgttga tggagactgg 40 <210> 784 <211> 20 <212> DNA <213> CNAG_02566 3 flanking region primer 2 <400> 784 actgcttctt caccctccac 20 <210> 785 <211> 18 <212> DNA <213> CNAG_02566 diagnostic screening primer, pairing with B79 <400> 785 atctgccgca acattcac 18 <210> 786 <211> 21 <212> DNA <213> CNAG_02566 Southern blot probe primer <400> 786 gttggtgctt catagtgact g 21 <210> 787 <211> 20 <212> DNA <213> NAT#219 STM primer <400> 787 ccctaaaacc ctacagcaat 20 <210> 788 <211> 24 <212> DNA <213> STM common primer <400> 788 gcatgccctg cccctaagaa ttcg 24 <210> 789 <211> 19 <212> DNA <213> CNAG_01626 5 flanking region primer 1 <400> 789 ggatgatgga atcgtatgc 19 <210> 790 <211> 38 <212> DNA <213> CNAG_01626 5 flanking region primer 2 <400> 790 tcactggccg tcgttttaca ttatccaccc tcggcttc 38 <210> 791 <211> 42 <212> DNA <213> CNAG_01626 3 flanking region primer 1 <400> 791 catggtcata gctgtttcct ggcgaacggt atgacaacta tg 42 <210> 792 <211> 22 <212> DNA <213> CNAG_01626 3 flanking region primer 2 <400> 792 ggacgaaaac attgctctca ac 22 <210> 793 <211> 19 <212> DNA <213> CNAG_01626 diagnostic screening primer, pairing with B79 <400> 793 tcagcagcag caggtaaac 19 <210> 794 <211> 18 <212> DNA <213> CNAG_01626 Southern blot probe primer <400> 794 cccagaccgt tttgaatg 18 <210> 795 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 795 ctttaaaggt ggtttgtg 18 <210> 796 <211> 24 <212> DNA <213> STM common primer <400> 796 gcatgccctg cccctaagaa ttcg 24 <210> 797 <211> 20 <212> DNA <213> CNAG_05940 5 flanking region primer 1 <400> 797 tcttgaccgt cgccagttac 20 <210> 798 <211> 39 <212> DNA <213> CNAG_05940 5 flanking region primer 2 <400> 798 tcactggccg tcgttttact ggctcaaagg caaagtcgg 39 <210> 799 <211> 40 <212> DNA <213> CNAG_05940 3 flanking region primer 1 <400> 799 catggtcata gctgtttcct gaggtgattt tgggagtcag 40 <210> 800 <211> 19 <212> DNA <213> CNAG_05940 3 flanking region primer 2 <400> 800 ggctgagaat gatgtgacg 19 <210> 801 <211> 19 <212> DNA <213> CNAG_05940 diagnostic screening primer, pairing with B79 <400> 801 ggaaagaaga cggatagcg 19 <210> 802 <211> 19 <212> DNA <213> CNAG_05940 Southern blot probe primer <400> 802 gtctccagga aagttcgtg 19 <210> 803 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 803 ctttaaaggt ggtttgtg 18 <210> 804 <211> 24 <212> DNA <213> STM common primer <400> 804 gcatgccctg cccctaagaa ttcg 24 <210> 805 <211> 22 <212> DNA <213> CNAG_04352 5 flanking region primer 1 <400> 805 gtgattatgt gaacccgtta cc 22 <210> 806 <211> 39 <212> DNA <213> CNAG_04352 5 flanking region primer 2 <400> 806 tcactggccg tcgttttact tttagaggtg atgactgcg 39 <210> 807 <211> 41 <212> DNA <213> CNAG_04352 3 flanking region primer 1 <400> 807 catggtcata gctgtttcct gtgctgctgg aggaagagaa c 41 <210> 808 <211> 19 <212> DNA <213> CNAG_04352 3 flanking region primer 2 <400> 808 agtgagtcgt tgatgtggc 19 <210> 809 <211> 19 <212> DNA <213> CNAG_04352 diagnostic screening primer, pairing with B79 <400> 809 atagtgaata cgggtcggg 19 <210> 810 <211> 19 <212> DNA <213> CNAG_04352 Southern blot probe primer <400> 810 gctgaaacag gaggcatag 19 <210> 811 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 811 ccagccagca ctccgatata 20 <210> 812 <211> 24 <212> DNA <213> STM common primer <400> 812 gcatgccctg cccctaagaa ttcg 24 <210> 813 <211> 18 <212> DNA <213> CNAG_03768 5 flanking region primer 1 <400> 813 aatgtccttg cggttgac 18 <210> 814 <211> 40 <212> DNA <213> CNAG_03768 5 flanking region primer 2 <400> 814 gctcactggc cgtcgtttta caatagtccc cgtcgttgtc 40 <210> 815 <211> 41 <212> DNA <213> CNAG_03768 3 flanking region primer 1 <400> 815 catggtcata gctgtttcct gcgatggatg tttggtggta g 41 <210> 816 <211> 19 <212> DNA <213> CNAG_03768 3 flanking region primer 2 <400> 816 aaaggggttt tggtggctc 19 <210> 817 <211> 20 <212> DNA <213> CNAG_03768 diagnostic screening primer, pairing with B79 <400> 817 tggatttggc agagaagaag 20 <210> 818 <211> 19 <212> DNA <213> CNAG_03768 Southern blot probe primer <400> 818 aagacacttg gctctccag 19 <210> 819 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 819 ccagccagca ctccgatata 20 <210> 820 <211> 24 <212> DNA <213> STM common primer <400> 820 gcatgccctg cccctaagaa ttcg 24 <210> 821 <211> 20 <212> DNA <213> CNAG_02322 5 flanking region primer 1 <400> 821 cgctcactca cctttcaaac 20 <210> 822 <211> 35 <212> DNA <213> CNAG_02322 5 flanking region primer 2 <400> 822 ctggccgtcg ttttacacat cggcttttac ggagc 35 <210> 823 <211> 37 <212> DNA <213> CNAG_02322 3 flanking region primer 1 <400> 823 gtcatagctg tttcctgctc ggttaccagt cactacg 37 <210> 824 <211> 20 <212> DNA <213> CNAG_02322 3 flanking region primer 2 <400> 824 ccaccaagat tcagagttcg 20 <210> 825 <211> 19 <212> DNA <213> CNAG_02322 diagnostic screening primer, pairing with B79 <400> 825 gagatttcgg tggatttcc 19 <210> 826 <211> 18 <212> DNA <213> CNAG_02322 Southern blot probe primer <400> 826 aagcaaccct cagcattg 18 <210> 827 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 827 ggtgttggat cggggtggat 20 <210> 828 <211> 24 <212> DNA <213> STM common primer <400> 828 gcatgccctg cccctaagaa ttcg 24 <210> 829 <211> 20 <212> DNA <213> CNAG_06818 5 flanking region primer 1 <400> 829 tgtattctgc ccacctccac 20 <210> 830 <211> 39 <212> DNA <213> CNAG_06818 5 flanking region primer 2 <400> 830 tcactggccg tcgttttacc ctctttgctg gtctctgag 39 <210> 831 <211> 41 <212> DNA <213> CNAG_06818 3 flanking region primer 1 <400> 831 catggtcata gctgtttcct gtctgctgct acatcaggtt c 41 <210> 832 <211> 19 <212> DNA <213> CNAG_06818 3 flanking region primer 2 <400> 832 aacctctgta tcgctgacg 19 <210> 833 <211> 22 <212> DNA <213> CNAG_06818 diagnostic screening primer, pairing with B79 <400> 833 gggtgaaaga caataggtaa gg 22 <210> 834 <211> 21 <212> DNA <213> CNAG_06818 Southern blot probe primer <400> 834 aaagtgaaga gaggaggtca g 21 <210> 835 <211> 20 <212> DNA <213> NAT#240 STM primer <400> 835 ggtgttggat cggggtggat 20 <210> 836 <211> 24 <212> DNA <213> STM common primer <400> 836 gcatgccctg cccctaagaa ttcg 24 <210> 837 <211> 19 <212> DNA <213> CNAG_07011 5 flanking region primer 1 <400> 837 ccaacaaata gcaaagccg 19 <210> 838 <211> 38 <212> DNA <213> CNAG_07011 5 flanking region primer 2 <400> 838 tcactggccg tcgttttaca caccaaagcc aagaacac 38 <210> 839 <211> 41 <212> DNA <213> CNAG_07011 3 flanking region primer 1 <400> 839 catggtcata gctgtttcct ggctaatgcc tgaaacagac c 41 <210> 840 <211> 19 <212> DNA <213> CNAG_07011 3 flanking region primer 2 <400> 840 tcggacaccc aatcaatac 19 <210> 841 <211> 18 <212> DNA <213> CNAG_07011 diagnostic screening primer, pairing with B79 <400> 841 ttgtgttgtt cggtgagg 18 <210> 842 <211> 20 <212> DNA <213> CNAG_07011 Southern blot probe primer <400> 842 tgggtgatac agaggaaaac 20 <210> 843 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 843 gagatctttc gggaggtctg gatt 24 <210> 844 <211> 24 <212> DNA <213> STM common primer <400> 844 gcatgccctg cccctaagaa ttcg 24 <210> 845 <211> 20 <212> DNA <213> CNAG_04036 5 flanking region primer 1 <400> 845 ccagttgaac gcttctaaac 20 <210> 846 <211> 39 <212> DNA <213> CNAG_04036 5 flanking region primer 2 <400> 846 tcactggccg tcgttttact gatggtgtgg gaacaatac 39 <210> 847 <211> 40 <212> DNA <213> CNAG_04036 3 flanking region primer 1 <400> 847 catggtcata gctgtttcct gttctccttg cctatgtgcc 40 <210> 848 <211> 20 <212> DNA <213> CNAG_04036 3 flanking region primer 2 <400> 848 cagcaactta ctttggaggc 20 <210> 849 <211> 19 <212> DNA <213> CNAG_04036 diagnostic screening primer, pairing with B79 <400> 849 atagacagca gcgggatac 19 <210> 850 <211> 18 <212> DNA <213> CNAG_04036 Southern blot probe primer <400> 850 tcacagatgc ctttctcg 18 <210> 851 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 851 gagatctttc gggaggtctg gatt 24 <210> 852 <211> 24 <212> DNA <213> STM common primer <400> 852 gcatgccctg cccctaagaa ttcg 24 <210> 853 <211> 19 <212> DNA <213> CNAG_07506 5 flanking region primer 1 <400> 853 cgtaatgaca ctggagagc 19 <210> 854 <211> 42 <212> DNA <213> CNAG_07506 5 flanking region primer 2 <400> 854 gctcactggc cgtcgtttta ctgttgccgt aggaaccgct tc 42 <210> 855 <211> 40 <212> DNA <213> CNAG_07506 3 flanking region primer 1 <400> 855 catggtcata gctgtttcct gtcccaaagc ctcttctcag 40 <210> 856 <211> 18 <212> DNA <213> CNAG_07506 3 flanking region primer 2 <400> 856 gtgagaccct gatagacc 18 <210> 857 <211> 19 <212> DNA <213> CNAG_07506 diagnostic screening primer, pairing with B79 <400> 857 gactcaaagg gagatgacg 19 <210> 858 <211> 19 <212> DNA <213> CNAG_07506 Southern blot probe primer <400> 858 gcttttgcct gctattgac 19 <210> 859 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 859 cgcccgccct cactatccac 20 <210> 860 <211> 24 <212> DNA <213> STM common primer <400> 860 gcatgccctg cccctaagaa ttcg 24 <210> 861 <211> 19 <212> DNA <213> CNAG_06223 5 flanking region primer 1 <400> 861 ggatgaaggg gaagacttg 19 <210> 862 <211> 38 <212> DNA <213> CNAG_06223 5 flanking region primer 2 <400> 862 tcactggccg tcgttttacc ttccgttcaa aatcgtcg 38 <210> 863 <211> 41 <212> DNA <213> CNAG_06223 3 flanking region primer 1 <400> 863 catggtcata gctgtttcct ggtctaccac caccatcaca c 41 <210> 864 <211> 20 <212> DNA <213> CNAG_06223 3 flanking region primer 2 <400> 864 gccgtccatt catacatctg 20 <210> 865 <211> 19 <212> DNA <213> CNAG_06223 diagnostic screening primer, pairing with B79 <400> 865 ggtgtcaata gttgggtgg 19 <210> 866 <211> 19 <212> DNA <213> CNAG_06223 Southern blot probe primer <400> 866 agagcagaag caaatgacc 19 <210> 867 <211> 20 <212> DNA <213> NAT#210 STM primer <400> 867 ctagagcccg ccacaacgct 20 <210> 868 <211> 24 <212> DNA <213> STM common primer <400> 868 gcatgccctg cccctaagaa ttcg 24 <210> 869 <211> 24 <212> DNA <213> CNAG_06283 5 flanking region primer 1 <400> 869 ccctttgtcc ttacccacat tttc 24 <210> 870 <211> 39 <212> DNA <213> CNAG_06283 5 flanking region primer 2 <400> 870 tcactggccg tcgttttact cgttcgggca cttcttgac 39 <210> 871 <211> 41 <212> DNA <213> CNAG_06283 3 flanking region primer 1 <400> 871 catggtcata gctgtttcct gcgagtaagg agacggttga g 41 <210> 872 <211> 22 <212> DNA <213> CNAG_06283 3 flanking region primer 2 <400> 872 ggttaggggg ataaaatagg ag 22 <210> 873 <211> 21 <212> DNA <213> CNAG_06283 diagnostic screening primer, pairing with B79 <400> 873 gagacattct ttctgacgag c 21 <210> 874 <211> 20 <212> DNA <213> CNAG_06283 Southern blot probe primer <400> 874 tggcttgaag ggtcttggtc 20 <210> 875 <211> 20 <212> DNA <213> NAT#234 STM primer <400> 875 ccagccagca ctccgatata 20 <210> 876 <211> 24 <212> DNA <213> STM common primer <400> 876 gcatgccctg cccctaagaa ttcg 24 <210> 877 <211> 22 <212> DNA <213> CNAG_04583 5 flanking region primer 1 <400> 877 gctcgctaca ttttcaggta tc 22 <210> 878 <211> 39 <212> DNA <213> CNAG_04583 5 flanking region primer 2 <400> 878 tcactggccg tcgttttacg agagggtcgg aaatagaag 39 <210> 879 <211> 41 <212> DNA <213> CNAG_04583 3 flanking region primer 1 <400> 879 catggtcata gctgtttcct ggggaaggag taatgcctga c 41 <210> 880 <211> 18 <212> DNA <213> CNAG_04583 3 flanking region primer 2 <400> 880 aaccgtgatg gagattgg 18 <210> 881 <211> 20 <212> DNA <213> CNAG_04583 diagnostic screening primer, pairing with B79 <400> 881 cggtgtgcga ttctaacttc 20 <210> 882 <211> 19 <212> DNA <213> CNAG_04583 Southern blot probe primer <400> 882 ggggactgtt ttgatgacc 19 <210> 883 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 883 ccatagcgat atctacccca atct 24 <210> 884 <211> 24 <212> DNA <213> STM common primer <400> 884 gcatgccctg cccctaagaa ttcg 24 <210> 885 <211> 19 <212> DNA <213> CNAG_03261 5 flanking region primer 1 <400> 885 ggcgataaag acgacgaac 19 <210> 886 <211> 35 <212> DNA <213> CNAG_03261 5 flanking region primer 2 <400> 886 ctggccgtcg ttttacggac tattcgtggc ttagc 35 <210> 887 <211> 35 <212> DNA <213> CNAG_03261 3 flanking region primer 1 <400> 887 gtcatagctg tttcctgtga gaccaatgtg ttgcc 35 <210> 888 <211> 24 <212> DNA <213> CNAG_03261 3 flanking region primer 2 <400> 888 cctcaacccc tttctgattt atcc 24 <210> 889 <211> 22 <212> DNA <213> CNAG_03261 diagnostic screening primer, pairing with B79 <400> 889 tagtcggtga tgagagttgc gg 22 <210> 890 <211> 20 <212> DNA <213> CNAG_03261 Southern blot probe primer <400> 890 gcaaatctgc ttctgggatg 20 <210> 891 <211> 19 <212> DNA <213> CNAG_04908 5 flanking region primer 1 <400> 891 gatgacgaat agcacggtc 19 <210> 892 <211> 37 <212> DNA <213> CNAG_04908 5 flanking region primer 2 <400> 892 tcactggccg tcgttttacg gatgggaaga ttgttgg 37 <210> 893 <211> 39 <212> DNA <213> CNAG_04908 3 flanking region primer 1 <400> 893 gatggtcata gctgtttcct gttgcttacg cctttgtcc 39 <210> 894 <211> 18 <212> DNA <213> CNAG_04908 3 flanking region primer 2 <400> 894 atggttgttc acgcacag 18 <210> 895 <211> 18 <212> DNA <213> CNAG_04908 diagnostic screening primer, pairing with B79 <400> 895 aaacaaaagc cgagggag 18 <210> 896 <211> 19 <212> DNA <213> CNAG_04908 Southern blot probe primer <400> 896 aacttcccac cccttcatc 19 <210> 897 <211> 24 <212> DNA <213> NAT#242 STM primer <400> 897 gtagcgatag gggtgtcgct ttag 24 <210> 898 <211> 24 <212> DNA <213> STM common primer <400> 898 gcatgccctg cccctaagaa ttcg 24 <210> 899 <211> 19 <212> DNA <213> CNAG_01317 5 flanking region primer 1 <400> 899 aagcaccaag aagcaagtg 19 <210> 900 <211> 37 <212> DNA <213> CNAG_01317 5 flanking region primer 2 <400> 900 tcactggccg tcgttttaca ttgacacctg ccgattc 37 <210> 901 <211> 40 <212> DNA <213> CNAG_01317 3 flanking region primer 1 <400> 901 catggtcata gctgtttcct gtactatcag cagaagcggc 40 <210> 902 <211> 20 <212> DNA <213> CNAG_01317 3 flanking region primer 2 <400> 902 aatgataggc ggaagtaagg 20 <210> 903 <211> 19 <212> DNA <213> CNAG_01317 diagnostic screening primer, pairing with B79 <400> 903 ccacctcgca gtatcaaac 19 <210> 904 <211> 20 <212> DNA <213> CNAG_01317 Southern blot probe primer <400> 904 agggtcaggg agatgatgtg 20 <210> 905 <211> 19 <212> DNA <213> CNAG_00559 5 flanking region primer 1 <400> 905 ggtgtttcca tacgaaccg 19 <210> 906 <211> 38 <212> DNA <213> CNAG_00559 5 flanking region primer 2 <400> 906 ctggccgtcg ttttacggag ccatttctta cttatgcg 38 <210> 907 <211> 35 <212> DNA <213> CNAG_00559 3 flanking region primer 1 <400> 907 gtcatagctg tttcctgaat cccaaggacc atctg 35 <210> 908 <211> 20 <212> DNA <213> CNAG_00559 3 flanking region primer 2 <400> 908 gctgagaagg agaggatagc 20 <210> 909 <211> 20 <212> DNA <213> CNAG_00559 diagnostic screening primer, pairing with B79 <400> 909 gtggttgaaa gtattaggcg 20 <210> 910 <211> 19 <212> DNA <213> CNAG_00559 Southern blot probe primer <400> 910 tctcttcgtt tgctacacg 19 <210> 911 <211> 24 <212> DNA <213> NAT#242 STM primer <400> 911 gtagcgatag gggtgtcgct ttag 24 <210> 912 <211> 24 <212> DNA <213> STM common primer <400> 912 gcatgccctg cccctaagaa ttcg 24 <210> 913 <211> 18 <212> DNA <213> CNAG_07940 5 flanking region primer 1 <400> 913 caaggtaatg attggggc 18 <210> 914 <211> 38 <212> DNA <213> CNAG_07940 5 flanking region primer 2 <400> 914 tcactggccg tcgttttacg caataatgga aggtctcg 38 <210> 915 <211> 39 <212> DNA <213> CNAG_07940 3 flanking region primer 1 <400> 915 catggtcata gctgtttcct gatcaagaat gacacgccg 39 <210> 916 <211> 19 <212> DNA <213> CNAG_07940 3 flanking region primer 2 <400> 916 aacttgtatg gtggtgccg 19 <210> 917 <211> 19 <212> DNA <213> CNAG_07940 diagnostic screening primer, pairing with B79 <400> 917 tcacttgctc ttctccctg 19 <210> 918 <211> 18 <212> DNA <213> CNAG_07940 Southern blot probe primer <400> 918 gtgacgaatg cgaaactc 18 <210> 919 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 919 atatggatgt ttttagcgag 20 <210> 920 <211> 24 <212> DNA <213> STM common primer <400> 920 gcatgccctg cccctaagaa ttcg 24 <210> 921 <211> 18 <212> DNA <213> CNAG_05535 5 flanking region primer 1 <400> 921 agcaacaacg gcatagtg 18 <210> 922 <211> 39 <212> DNA <213> CNAG_05535 5 flanking region primer 2 <400> 922 tcactggccg tcgttttaca cgacagtgag agtcctgac 39 <210> 923 <211> 43 <212> DNA <213> CNAG_05535 3 flanking region primer 1 <400> 923 catggtcata gctgtttcct gctggagcga ggaaatagta atg 43 <210> 924 <211> 18 <212> DNA <213> CNAG_05535 3 flanking region primer 2 <400> 924 acacaaagcg agtggttg 18 <210> 925 <211> 19 <212> DNA <213> CNAG_05535 diagnostic screening primer, pairing with B79 <400> 925 ggcagtgtca gatgaaagc 19 <210> 926 <211> 19 <212> DNA <213> CNAG_05535 Southern blot probe primer <400> 926 catcggctac atcttcctc 19 <210> 927 <211> 21 <212> DNA <213> CNAG_02516 5 flanking region primer 1 <400> 927 gcatcacacg aacgattcta c 21 <210> 928 <211> 38 <212> DNA <213> CNAG_02516 5 flanking region primer 2 <400> 928 tcactggccg tcgttttaca ttgttggcgg tggaaacg 38 <210> 929 <211> 41 <212> DNA <213> CNAG_02516 3 flanking region primer 1 <400> 929 catggtcata gctgtttcct gaggtttggg agcagtaagg c 41 <210> 930 <211> 19 <212> DNA <213> CNAG_02516 3 flanking region primer 2 <400> 930 gatgtttgtg accgattcg 19 <210> 931 <211> 19 <212> DNA <213> CNAG_02516 diagnostic screening primer, pairing with B79 <400> 931 gcgaaggttt accgataag 19 <210> 932 <211> 20 <212> DNA <213> CNAG_02516 Southern blot probe primer <400> 932 tatggggtta ggagcgttgg 20 <210> 933 <211> 20 <212> DNA <213> NAT#210 STM primer <400> 933 ctagagcccg ccacaacgct 20 <210> 934 <211> 24 <212> DNA <213> STM common primer <400> 934 gcatgccctg cccctaagaa ttcg 24 <210> 935 <211> 19 <212> DNA <213> CNAG_04226 5 flanking region primer 1 <400> 935 gccctttttg gtatgacag 19 <210> 936 <211> 38 <212> DNA <213> CNAG_04226 5 flanking region primer 2 <400> 936 tcactggccg tcgttttact gtgagggatt ccttgttc 38 <210> 937 <211> 39 <212> DNA <213> CNAG_04226 3 flanking region primer 1 <400> 937 catggtcata gctgtttcct ggttggaaac aagcagtcg 39 <210> 938 <211> 20 <212> DNA <213> CNAG_04226 3 flanking region primer 2 <400> 938 tcagaagaat caagaggtcg 20 <210> 939 <211> 19 <212> DNA <213> CNAG_04226 diagnostic screening primer, pairing with B79 <400> 939 acttacattg ccttcctcg 19 <210> 940 <211> 19 <212> DNA <213> CNAG_04226 Southern blot probe primer <400> 940 atacatcagc ctgtcctgc 19 <210> 941 <211> 19 <212> DNA <213> CNAG_05170 5 flanking region primer 1 <400> 941 tgccttcgct ctgtaactc 19 <210> 942 <211> 34 <212> DNA <213> CNAG_05170 5 flanking region primer 2 <400> 942 ctggccgtcg ttttaccctt tttgaggctc gctg 34 <210> 943 <211> 36 <212> DNA <213> CNAG_05170 3 flanking region primer 1 <400> 943 gtcatagctg tttcctgaaa tgaaggagtg tgcggg 36 <210> 944 <211> 20 <212> DNA <213> CNAG_05170 3 flanking region primer 2 <400> 944 aataggacct caacaccagg 20 <210> 945 <211> 18 <212> DNA <213> CNAG_05170 diagnostic screening primer, pairing with B79 <400> 945 aacaagaagc ccctttcg 18 <210> 946 <211> 18 <212> DNA <213> CNAG_05170 Southern blot probe primer <400> 946 agtttttgga cccgagtg 18 <210> 947 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 947 ctttaaaggt ggtttgtg 18 <210> 948 <211> 24 <212> DNA <213> STM common primer <400> 948 gcatgccctg cccctaagaa ttcg 24 <210> 949 <211> 20 <212> DNA <213> CNAG_03902 5 flanking region primer 1 <400> 949 gcaaagtcaa gaaggtcagg 20 <210> 950 <211> 35 <212> DNA <213> CNAG_03902 5 flanking region primer 2 <400> 950 ctggccgtcg ttttacttct tctttggctc gtccg 35 <210> 951 <211> 39 <212> DNA <213> CNAG_03902 3 flanking region primer 1 <400> 951 gtcatagctg tttcctgggg agatgaaatg tgcgtttag 39 <210> 952 <211> 22 <212> DNA <213> CNAG_03902 3 flanking region primer 2 <400> 952 atgctttcgc cgtcatctcg tc 22 <210> 953 <211> 20 <212> DNA <213> CNAG_03902 diagnostic screening primer, pairing with B79 <400> 953 tgattacccc tgtcaaactc 20 <210> 954 <211> 19 <212> DNA <213> CNAG_03902 Southern blot probe primer <400> 954 cagaatcggg aagtaatgg 19 <210> 955 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 955 ctttaaaggt ggtttgtg 18 <210> 956 <211> 24 <212> DNA <213> STM common primer <400> 956 gcatgccctg cccctaagaa ttcg 24 <210> 957 <211> 20 <212> DNA <213> CNAG_06097 5 flanking region primer 1 <400> 957 cagtccgaag tttacaatcg 20 <210> 958 <211> 36 <212> DNA <213> CNAG_06097 5 flanking region primer 2 <400> 958 ctggccgtcg ttttactttc gtcgtgtgga gttggc 36 <210> 959 <211> 37 <212> DNA <213> CNAG_06097 3 flanking region primer 1 <400> 959 gtcatagctg tttcctgaga ttggagggtt tgagggc 37 <210> 960 <211> 19 <212> DNA <213> CNAG_06097 3 flanking region primer 2 <400> 960 tccaccgaag aaactgaag 19 <210> 961 <211> 20 <212> DNA <213> CNAG_06097 diagnostic screening primer, pairing with B79 <400> 961 aggtttgaga ggtttggaag 20 <210> 962 <211> 18 <212> DNA <213> CNAG_06097 Southern blot probe primer <400> 962 attatcggag cgttgtgg 18 <210> 963 <211> 20 <212> DNA <213> CNAG_03431 5 flanking region primer 1 <400> 963 aagcactctg tagcctgttg 20 <210> 964 <211> 39 <212> DNA <213> CNAG_03431 5 flanking region primer 2 <400> 964 gctcactggc cgtcgtttta ccttgaagat gatgcgtgc 39 <210> 965 <211> 40 <212> DNA <213> CNAG_03431 3 flanking region primer 1 <400> 965 catggtcata gctgtttcct gcgctcatag agaatgctgg 40 <210> 966 <211> 19 <212> DNA <213> CNAG_03431 3 flanking region primer 2 <400> 966 taggagtgtc tgccaaagc 19 <210> 967 <211> 19 <212> DNA <213> CNAG_03431 diagnostic screening primer, pairing with B79 <400> 967 tcatcctcat cccgaatac 19 <210> 968 <211> 19 <212> DNA <213> CNAG_03431 Southern blot probe primer <400> 968 actcttcgtg agacgcatc 19 <210> 969 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 969 accgacagct cgaacaagca agag 24 <210> 970 <211> 24 <212> DNA <213> STM common primer <400> 970 gcatgccctg cccctaagaa ttcg 24 <210> 971 <211> 21 <212> DNA <213> CNAG_00830 5 flanking region primer 1 <400> 971 ccgactaata ggtgtgctgt c 21 <210> 972 <211> 41 <212> DNA <213> CNAG_00830 5 flanking region primer 2 <400> 972 gctcactggc cgtcgtttta cgagatgaga tgagttgcgt g 41 <210> 973 <211> 41 <212> DNA <213> CNAG_00830 3 flanking region primer 1 <400> 973 catggtcata gctgtttcct gttgtgtccc catcactacc g 41 <210> 974 <211> 19 <212> DNA <213> CNAG_00830 3 flanking region primer 2 <400> 974 catccaaaga agcgaaacg 19 <210> 975 <211> 24 <212> DNA <213> CNAG_00830 diagnostic screening primer, pairing with B79 <400> 975 gaagagaagt agtggataga ctcg 24 <210> 976 <211> 19 <212> DNA <213> CNAG_00830 Southern blot probe primer <400> 976 tgatgccgta aatcgtagg 19 <210> 977 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 977 atttcgatgt atatatgtag c 21 <210> 978 <211> 24 <212> DNA <213> STM common primer <400> 978 gcatgccctg cccctaagaa ttcg 24 <210> 979 <211> 20 <212> DNA <213> CNAG_07443 5 flanking region primer 1 <400> 979 atctcttctc tcggcgtgtc 20 <210> 980 <211> 40 <212> DNA <213> CNAG_07443 5 flanking region primer 2 <400> 980 gctcactggc cgtcgtttta cacaggaaaa ggaggagtcg 40 <210> 981 <211> 40 <212> DNA <213> CNAG_07443 3 flanking region primer 1 <400> 981 catggtcata gctgtttcct gaagtctttc cgtctgaccc 40 <210> 982 <211> 18 <212> DNA <213> CNAG_07443 3 flanking region primer 2 <400> 982 tcgcatccct tgtattcg 18 <210> 983 <211> 19 <212> DNA <213> CNAG_07443 diagnostic screening primer, pairing with B79 <400> 983 tcaataatca gccgcctac 19 <210> 984 <211> 19 <212> DNA <213> CNAG_07443 Southern blot probe primer <400> 984 ttagattccg cataggtcc 19 <210> 985 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 985 acacctacat caaaccctcc c 21 <210> 986 <211> 24 <212> DNA <213> STM common primer <400> 986 gcatgccctg cccctaagaa ttcg 24 <210> 987 <211> 19 <212> DNA <213> CNAG_05186 5 flanking region primer 1 <400> 987 acaacggtca agaagggtc 19 <210> 988 <211> 39 <212> DNA <213> CNAG_05186 5 flanking region primer 2 <400> 988 gctcactggc cgtcgtttta caagtaaagg aacgcacgg 39 <210> 989 <211> 41 <212> DNA <213> CNAG_05186 3 flanking region primer 1 <400> 989 catggtcata gctgtttcct gcgcttgaga cgggagtaat c 41 <210> 990 <211> 20 <212> DNA <213> CNAG_05186 3 flanking region primer 2 <400> 990 gaggtaagga tgattggacc 20 <210> 991 <211> 23 <212> DNA <213> CNAG_05186 diagnostic screening primer, pairing with B79 <400> 991 gtgtgaagtg agtagtaacg agc 23 <210> 992 <211> 22 <212> DNA <213> CNAG_05186 Southern blot probe primer <400> 992 ttccctatct ctcttcatac cc 22 <210> 993 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 993 cgcccgccct cactatccac 20 <210> 994 <211> 24 <212> DNA <213> STM common primer <400> 994 gcatgccctg cccctaagaa ttcg 24 <210> 995 <211> 20 <212> DNA <213> CNAG_03561 5 flanking region primer 1 <400> 995 tgggcgagac gagatagaac 20 <210> 996 <211> 36 <212> DNA <213> CNAG_03561 5 flanking region primer 2 <400> 996 ctggccgtcg ttttacggat aaagatgatt cgggtc 36 <210> 997 <211> 36 <212> DNA <213> CNAG_03561 3 flanking region primer 1 <400> 997 gtcatagctg tttcctgtga tggctgattt cccgtg 36 <210> 998 <211> 36 <212> DNA <213> CNAG_03561 3 flanking region primer 2 <400> 998 gtcatagctg tttcctgtga tggctgattt cccgtg 36 <210> 999 <211> 20 <212> DNA <213> CNAG_03561 diagnostic screening primer, pairing with B79 <400> 999 aagaaagacg gtagcggtgg 20 <210> 1000 <211> 20 <212> DNA <213> CNAG_03561 Southern blot probe primer <400> 1000 ccacctcttc atccaatagc 20 <210> 1001 <211> 20 <212> DNA <213> NAT#43 STM primer <400> 1001 ccagctacca atcacgctac 20 <210> 1002 <211> 24 <212> DNA <213> STM common primer <400> 1002 gcatgccctg cccctaagaa ttcg 24 <210> 1003 <211> 19 <212> DNA <213> CNAG_04878 5 flanking region primer 1 <400> 1003 aagtccgtgc tggctaatc 19 <210> 1004 <211> 35 <212> DNA <213> CNAG_04878 5 flanking region primer 2 <400> 1004 ctggccgtcg ttttaccttt tcgctgcctt attcg 35 <210> 1005 <211> 41 <212> DNA <213> CNAG_04878 3 flanking region primer 1 <400> 1005 gtcatagctg tttcctgaaa acagggcata cggtgtcgtg g 41 <210> 1006 <211> 20 <212> DNA <213> CNAG_04878 3 flanking region primer 2 <400> 1006 gctgaccatc tcttccctac 20 <210> 1007 <211> 22 <212> DNA <213> CNAG_04878 diagnostic screening primer, pairing with B79 <400> 1007 ccttgtttgt gtctaccgta tc 22 <210> 1008 <211> 19 <212> DNA <213> CNAG_04878 Southern blot probe primer <400> 1008 agtgggaatc gtcgttgac 19 <210> 1009 <211> 20 <212> DNA <213> NAT#116 STM primer <400> 1009 gcacccaaga gctccatctc 20 <210> 1010 <211> 24 <212> DNA <213> STM common primer <400> 1010 gcatgccctg cccctaagaa ttcg 24 <210> 1011 <211> 19 <212> DNA <213> CNAG_03279 5 flanking region primer 1 <400> 1011 cgtttccagt tatcgcttc 19 <210> 1012 <211> 41 <212> DNA <213> CNAG_03279 5 flanking region primer 2 <400> 1012 gctcactggc cgtcgtttta cgcaatgacc cactttttga c 41 <210> 1013 <211> 41 <212> DNA <213> CNAG_03279 3 flanking region primer 1 <400> 1013 catggtcata gctgtttcct ggctttgcct gatgagtttt g 41 <210> 1014 <211> 19 <212> DNA <213> CNAG_03279 3 flanking region primer 2 <400> 1014 gtaacccact cggaagttg 19 <210> 1015 <211> 19 <212> DNA <213> CNAG_03279 diagnostic screening primer, pairing with B79 <400> 1015 ttgtctgtga ggtggaagg 19 <210> 1016 <211> 20 <212> DNA <213> CNAG_03279 Southern blot probe primer <400> 1016 caggctcgtt tgtagccttc 20 <210> 1017 <211> 24 <212> DNA <213> NAT#122 STM primer <400> 1017 acagctccaa acctcgctaa acag 24 <210> 1018 <211> 24 <212> DNA <213> STM common primer <400> 1018 gcatgccctg cccctaagaa ttcg 24 <210> 1019 <211> 18 <212> DNA <213> CNAG_06921 5 flanking region primer 1 <400> 1019 gacaaagcaa caacagcg 18 <210> 1020 <211> 37 <212> DNA <213> CNAG_06921 5 flanking region primer 2 <400> 1020 tcactggccg tcgttttact cggatgacag atggttg 37 <210> 1021 <211> 39 <212> DNA <213> CNAG_06921 3 flanking region primer 1 <400> 1021 catggtcata gctgtttcct gaaagattgg gcgtctctg 39 <210> 1022 <211> 18 <212> DNA <213> CNAG_06921 3 flanking region primer 2 <400> 1022 atcatcagcg acatccag 18 <210> 1023 <211> 18 <212> DNA <213> CNAG_06921 diagnostic screening primer, pairing with B79 <400> 1023 aaagaacata cacgccgc 18 <210> 1024 <211> 18 <212> DNA <213> CNAG_06921 Southern blot probe primer <400> 1024 accatctgga tttctgcc 18 <210> 1025 <211> 21 <212> DNA <213> NAT#159 STM primer <400> 1025 acgcaccaga cacacaacca g 21 <210> 1026 <211> 24 <212> DNA <213> STM common primer <400> 1026 gcatgccctg cccctaagaa ttcg 24 <210> 1027 <211> 20 <212> DNA <213> CNAG_02555 5 flanking region primer 1 <400> 1027 agaggaacac ggtatggtcg 20 <210> 1028 <211> 40 <212> DNA <213> CNAG_02555 5 flanking region primer 2 <400> 1028 gctcactggc cgtcgtttta ccaacaacgg tgaaccaaag 40 <210> 1029 <211> 41 <212> DNA <213> CNAG_02555 3 flanking region primer 1 <400> 1029 catggtcata gctgtttcct gcagagtgga ggtagaaacg g 41 <210> 1030 <211> 18 <212> DNA <213> CNAG_02555 3 flanking region primer 2 <400> 1030 tctgaaatgc tggctgtc 18 <210> 1031 <211> 20 <212> DNA <213> CNAG_02555 diagnostic screening primer, pairing with B79 <400> 1031 cccctcacaa cttgctaatg 20 <210> 1032 <211> 20 <212> DNA <213> CNAG_02555 Southern blot probe primer <400> 1032 ccgatgtcga cgtatgattg 20 <210> 1033 <211> 18 <212> DNA <213> NAT#270 STM primer <400> 1033 ggtggtttgt gtgggtag 18 <210> 1034 <211> 24 <212> DNA <213> STM common primer <400> 1034 gcatgccctg cccctaagaa ttcg 24 <210> 1035 <211> 19 <212> DNA <213> CNAG_03710 5 flanking region primer 1 <400> 1035 aagacaagaa gcagcagcg 19 <210> 1036 <211> 40 <212> DNA <213> CNAG_03710 5 flanking region primer 2 <400> 1036 gctcactggc cgtcgtttta ctatcattcc catccgtcgc 40 <210> 1037 <211> 42 <212> DNA <213> CNAG_03710 3 flanking region primer 1 <400> 1037 catggtcata gctgtttcct gcgctcaata acatcgtcag ac 42 <210> 1038 <211> 20 <212> DNA <213> CNAG_03710 3 flanking region primer 2 <400> 1038 ccagccttca tctctcttcc 20 <210> 1039 <211> 20 <212> DNA <213> CNAG_03710 diagnostic screening primer, pairing with B79 <400> 1039 cagaactgat aactggctgc 20 <210> 1040 <211> 20 <212> DNA <213> CNAG_03710 Southern blot probe primer <400> 1040 tcctctgcac caagactgtg 20 <210> 1041 <211> 24 <212> DNA <213> NAT#290 STM primer <400> 1041 accgacagct cgaacaagca agag 24 <210> 1042 <211> 24 <212> DNA <213> STM common primer <400> 1042 gcatgccctg cccctaagaa ttcg 24 <210> 1043 <211> 20 <212> DNA <213> CNAG_03183 5 flanking region primer 1 <400> 1043 ctctatgagg gcaaagtcag 20 <210> 1044 <211> 40 <212> DNA <213> CNAG_03183 5 flanking region primer 2 <400> 1044 gctcactggc cgtcgtttta ctgttgttgc tgttggctcg 40 <210> 1045 <211> 40 <212> DNA <213> CNAG_03183 3 flanking region primer 1 <400> 1045 catggtcata gctgtttcct gttcttccaa tcccagccac 40 <210> 1046 <211> 19 <212> DNA <213> CNAG_03183 3 flanking region primer 2 <400> 1046 tacgcttcac agattacgc 19 <210> 1047 <211> 18 <212> DNA <213> CNAG_03183 diagnostic screening primer, pairing with B79 <400> 1047 tatacgtcca agacatcc 18 <210> 1048 <211> 20 <212> DNA <213> CNAG_03183 Southern blot probe primer <400> 1048 ctgattccga ggcttactcg 20 <210> 1049 <211> 21 <212> DNA <213> NAT#292 STM primer <400> 1049 atttcgatgt atatatgtag c 21 <210> 1050 <211> 24 <212> DNA <213> STM common primer <400> 1050 gcatgccctg cccctaagaa ttcg 24 <210> 1051 <211> 20 <212> DNA <213> CNAG_04637 5 flanking region primer 1 <400> 1051 ccttcaagcc aaaatgagac 20 <210> 1052 <211> 40 <212> DNA <213> CNAG_04637 5 flanking region primer 2 <400> 1052 gctcactggc cgtcgtttta cgagatggag agaagcaacg 40 <210> 1053 <211> 43 <212> DNA <213> CNAG_04637 3 flanking region primer 1 <400> 1053 catggtcata gctgtttcct gggttgacgg atttactgaa gtc 43 <210> 1054 <211> 20 <212> DNA <213> CNAG_04637 3 flanking region primer 2 <400> 1054 tggaagatga ctcactcggg 20 <210> 1055 <211> 19 <212> DNA <213> CNAG_04637 diagnostic screening primer, pairing with B79 <400> 1055 tccatcaaag atacacccg 19 <210> 1056 <211> 20 <212> DNA <213> CNAG_04637 Southern blot probe primer <400> 1056 ttacatcttg aggcttagcg 20 <210> 1057 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 1057 cgcccgccct cactatccac 20 <210> 1058 <211> 24 <212> DNA <213> STM common primer <400> 1058 gcatgccctg cccctaagaa ttcg 24 <210> 1059 <211> 19 <212> DNA <213> CNAG_04398 5 flanking region primer 1 <400> 1059 agtctggaga aaaggggtc 19 <210> 1060 <211> 34 <212> DNA <213> CNAG_04398 5 flanking region primer 2 <400> 1060 ctggccgtcg ttttacgcaa acacacattc tcgg 34 <210> 1061 <211> 36 <212> DNA <213> CNAG_04398 3 flanking region primer 1 <400> 1061 gtcatagctg tttcctgttg gctaatgtag ggttgg 36 <210> 1062 <211> 18 <212> DNA <213> CNAG_04398 3 flanking region primer 2 <400> 1062 ccccttccca tttttcac 18 <210> 1063 <211> 22 <212> DNA <213> CNAG_04398 diagnostic screening primer, pairing with B79 <400> 1063 gccactttct gatttccgta tc 22 <210> 1064 <211> 18 <212> DNA <213> CNAG_04398 Southern blot probe primer <400> 1064 tatttgctgc ctgaccac 18 <210> 1065 <211> 20 <212> DNA <213> NAT#225 STM primer <400> 1065 ccatagaact agctaaagca 20 <210> 1066 <211> 24 <212> DNA <213> STM common primer <400> 1066 gcatgccctg cccctaagaa ttcg 24 <210> 1067 <211> 20 <212> DNA <213> CNAG_01454 5 flanking region primer 1 <400> 1067 ggaagtttgg aggaggatag 20 <210> 1068 <211> 37 <212> DNA <213> CNAG_01454 5 flanking region primer 2 <400> 1068 tcactggccg tcgttttacc agattcgtgg tcaaagc 37 <210> 1069 <211> 40 <212> DNA <213> CNAG_01454 3 flanking region primer 1 <400> 1069 catggtcata gctgtttcct ggccttgaat gatgaccagc 40 <210> 1070 <211> 18 <212> DNA <213> CNAG_01454 3 flanking region primer 2 <400> 1070 cggatttgag atgatgcc 18 <210> 1071 <211> 22 <212> DNA <213> CNAG_01454 diagnostic screening primer, pairing with B79 <400> 1071 gttagaggag tcattctttg gc 22 <210> 1072 <211> 20 <212> DNA <213> CNAG_01454 Southern blot probe primer <400> 1072 caaaaggtca aggaaaggtg 20 <210> 1073 <211> 24 <212> DNA <213> NAT#58 STM primer <400> 1073 cgcaaaatca ctagccctat agcg 24 <210> 1074 <211> 24 <212> DNA <213> STM common primer <400> 1074 gcatgccctg cccctaagaa ttcg 24 <210> 1075 <211> 18 <212> DNA <213> CNAG_04268 5 flanking region primer 1 <400> 1075 tcgtctttgt gtggttgc 18 <210> 1076 <211> 38 <212> DNA <213> CNAG_04268 5 flanking region primer 2 <400> 1076 tcactggccg tcgttttaca cttatggcga cttacggg 38 <210> 1077 <211> 40 <212> DNA <213> CNAG_04268 3 flanking region primer 1 <400> 1077 catggtcata gctgtttcct ggcaatgtct gtatcaccgc 40 <210> 1078 <211> 19 <212> DNA <213> CNAG_04268 3 flanking region primer 2 <400> 1078 cgtccgcctg tattttatc 19 <210> 1079 <211> 18 <212> DNA <213> CNAG_04268 diagnostic screening primer, pairing with B79 <400> 1079 atgatgctgt tttgcccc 18 <210> 1080 <211> 18 <212> DNA <213> CNAG_04268 Southern blot probe primer <400> 1080 ttctttccac atcggtcc 18 <210> 1081 <211> 24 <212> DNA <213> NAT#102 STM primer <400> 1081 ccatagcgat atctacccca atct 24 <210> 1082 <211> 24 <212> DNA <213> STM common primer <400> 1082 gcatgccctg cccctaagaa ttcg 24 <210> 1083 <211> 22 <212> DNA <213> CNAG_00031 5 flanking region primer 1 <400> 1083 atccaatcat cggcttccca gc 22 <210> 1084 <211> 35 <212> DNA <213> CNAG_00031 5 flanking region primer 2 <400> 1084 ctggccgtcg ttttacactt gaactcccat ctgcg 35 <210> 1085 <211> 37 <212> DNA <213> CNAG_00031 3 flanking region primer 1 <400> 1085 gtcatagctg tttcctggaa atgcgatgaa tgggttg 37 <210> 1086 <211> 21 <212> DNA <213> CNAG_00031 3 flanking region primer 2 <400> 1086 cctcttgccc taatgagata g 21 <210> 1087 <211> 18 <212> DNA <213> CNAG_00031 diagnostic screening primer, pairing with B79 <400> 1087 ccgtgttcgc aatctatg 18 <210> 1088 <211> 19 <212> DNA <213> CNAG_00031 Southern blot probe primer <400> 1088 gaatcagcat ttgggttcg 19 <210> 1089 <211> 20 <212> DNA <213> NAT#116 STM primer <400> 1089 gcacccaaga gctccatctc 20 <210> 1090 <211> 24 <212> DNA <213> STM common primer <400> 1090 gcatgccctg cccctaagaa ttcg 24 <210> 1091 <211> 19 <212> DNA <213> CNAG_07922 5 flanking region primer 1 <400> 1091 cctcttccgc tattttgtg 19 <210> 1092 <211> 39 <212> DNA <213> CNAG_07922 5 flanking region primer 2 <400> 1092 gctcactggc cgtcgtttta ccattgctgt tgtgggaac 39 <210> 1093 <211> 41 <212> DNA <213> CNAG_07922 3 flanking region primer 1 <400> 1093 catggtcata gctgtttcct gaagtggagt tggaagaggc g 41 <210> 1094 <211> 22 <212> DNA <213> CNAG_07922 3 flanking region primer 2 <400> 1094 caactatcca atacctgttc cg 22 <210> 1095 <211> 19 <212> DNA <213> CNAG_07922 diagnostic screening primer, pairing with B79 <400> 1095 cacggcttgt cttgaagtc 19 <210> 1096 <211> 18 <212> DNA <213> CNAG_07922 Southern blot probe primer <400> 1096 ttgacggttt tggcacag 18 <210> 1097 <211> 20 <212> DNA <213> NAT#116 STM primer <400> 1097 gcacccaaga gctccatctc 20 <210> 1098 <211> 24 <212> DNA <213> STM common primer <400> 1098 gcatgccctg cccctaagaa ttcg 24 <210> 1099 <211> 19 <212> DNA <213> CNAG_04841 5 flanking region primer 1 <400> 1099 tgaggtgttc cgtgtattg 19 <210> 1100 <211> 35 <212> DNA <213> CNAG_04841 5 flanking region primer 2 <400> 1100 ctggccgtcg ttttacctct tgactggtaa tgggg 35 <210> 1101 <211> 35 <212> DNA <213> CNAG_04841 3 flanking region primer 1 <400> 1101 gtcatagctg tttcctgatt cccaaggtga cgaag 35 <210> 1102 <211> 20 <212> DNA <213> CNAG_04841 3 flanking region primer 2 <400> 1102 tctccaataa gttcaggcac 20 <210> 1103 <211> 22 <212> DNA <213> CNAG_04841 diagnostic screening primer, pairing with B79 <400> 1103 taaagacgga ggtgggagta gg 22 <210> 1104 <211> 18 <212> DNA <213> CNAG_04841 Southern blot probe primer <400> 1104 tgagattggg aaagagcg 18 <210> 1105 <211> 24 <212> DNA <213> NAT#119 STM primer <400> 1105 ctccccacat aaagagagct aaac 24 <210> 1106 <211> 24 <212> DNA <213> STM common primer <400> 1106 gcatgccctg cccctaagaa ttcg 24 <210> 1107 <211> 20 <212> DNA <213> CNAG_01948 5 flanking region primer 1 <400> 1107 cttggtaaat ccgtattccg 20 <210> 1108 <211> 35 <212> DNA <213> CNAG_01948 5 flanking region primer 2 <400> 1108 ctggccgtcg ttttaccctt tcgcttcaac ctacg 35 <210> 1109 <211> 35 <212> DNA <213> CNAG_01948 3 flanking region primer 1 <400> 1109 gtcatagctg tttcctgtga tttggcgact ctgag 35 <210> 1110 <211> 20 <212> DNA <213> CNAG_01948 3 flanking region primer 2 <400> 1110 tgtgtttctc atcgtaagcg 20 <210> 1111 <211> 22 <212> DNA <213> CNAG_01948 diagnostic screening primer, pairing with B79 <400> 1111 ttgacttccc aggttacact ac 22 <210> 1112 <211> 19 <212> DNA <213> CNAG_01948 Southern blot probe primer <400> 1112 cattgggact cttgtaggg 19 <210> 1113 <211> 24 <212> DNA <213> NAT#119 STM primer <400> 1113 ctccccacat aaagagagct aaac 24 <210> 1114 <211> 24 <212> DNA <213> STM common primer <400> 1114 gcatgccctg cccctaagaa ttcg 24 <210> 1115 <211> 19 <212> DNA <213> CNAG_07435 5 flanking region primer 1 <400> 1115 caaccttcaa agtcttgcc 19 <210> 1116 <211> 39 <212> DNA <213> CNAG_07435 5 flanking region primer 2 <400> 1116 tcactggccg tcgttttaca acgaggagtc atactgtgc 39 <210> 1117 <211> 41 <212> DNA <213> CNAG_07435 3 flanking region primer 1 <400> 1117 catggtcata gctgtttcct gaaaggcgtc aaagggcgaa g 41 <210> 1118 <211> 20 <212> DNA <213> CNAG_07435 3 flanking region primer 2 <400> 1118 ggtttggagc gtattgattg 20 <210> 1119 <211> 19 <212> DNA <213> CNAG_07435 diagnostic screening primer, pairing with B79 <400> 1119 aagagcggca ttgaatcgg 19 <210> 1120 <211> 20 <212> DNA <213> CNAG_07435 Southern blot probe primer <400> 1120 gcgtcacttt ggacatactc 20 <210> 1121 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 1121 ctatcgacca accaacacag 20 <210> 1122 <211> 24 <212> DNA <213> STM common primer <400> 1122 gcatgccctg cccctaagaa ttcg 24 <210> 1123 <211> 22 <212> DNA <213> CNAG_00505 5 flanking region primer 1 <400> 1123 tttcaggcac caacaggcac ag 22 <210> 1124 <211> 38 <212> DNA <213> CNAG_00505 5 flanking region primer 2 <400> 1124 ctggccgtcg ttttacttcc gatggtgccc ttgagtag 38 <210> 1125 <211> 38 <212> DNA <213> CNAG_00505 3 flanking region primer 1 <400> 1125 gtcatagctg tttcctgggt cttccaaatg tcacagtc 38 <210> 1126 <211> 19 <212> DNA <213> CNAG_00505 3 flanking region primer 2 <400> 1126 aaagaagaga cgcctcacg 19 <210> 1127 <211> 20 <212> DNA <213> CNAG_00505 diagnostic screening primer, pairing with B79 <400> 1127 tgatagacac cacggcagtc 20 <210> 1128 <211> 20 <212> DNA <213> CNAG_00505 Southern blot probe primer <400> 1128 cgaggattgt caaatgaagg 20 <210> 1129 <211> 24 <212> DNA <213> NAT#125 STM primer <400> 1129 cgctacagcc agcgcgcgca agcg 24 <210> 1130 <211> 24 <212> DNA <213> STM common primer <400> 1130 gcatgccctg cccctaagaa ttcg 24 <210> 1131 <211> 18 <212> DNA <213> CNAG_03115 5 flanking region primer 1 <400> 1131 cgaaggaagt aaaacggg 18 <210> 1132 <211> 37 <212> DNA <213> CNAG_03115 5 flanking region primer 2 <400> 1132 ctggccgtcg ttttacaagc actcaccact ctcctac 37 <210> 1133 <211> 36 <212> DNA <213> CNAG_03115 3 flanking region primer 1 <400> 1133 gtcatagctg tttcctgtgc ttggatagac gacgag 36 <210> 1134 <211> 20 <212> DNA <213> CNAG_03115 3 flanking region primer 2 <400> 1134 tcaatcaaag cgttcggggg 20 <210> 1135 <211> 20 <212> DNA <213> CNAG_03115 diagnostic screening primer, pairing with B79 <400> 1135 aatccgactt ctcagccctg 20 <210> 1136 <211> 20 <212> DNA <213> CNAG_03115 Southern blot probe primer <400> 1136 atctgaggga agggctacag 20 <210> 1137 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 1137 caccaactcc ccatctccat 20 <210> 1138 <211> 24 <212> DNA <213> STM common primer <400> 1138 gcatgccctg cccctaagaa ttcg 24 <210> 1139 <211> 19 <212> DNA <213> CNAG_04807 5 flanking region primer 1 <400> 1139 gcgtttcctc gtgaacttc 19 <210> 1140 <211> 37 <212> DNA <213> CNAG_04807 5 flanking region primer 2 <400> 1140 ctggccgtcg ttttacggaa gaagggttga gacactc 37 <210> 1141 <211> 36 <212> DNA <213> CNAG_04807 3 flanking region primer 1 <400> 1141 gtcatagctg tttcctgggc aagcataatg gcaaag 36 <210> 1142 <211> 18 <212> DNA <213> CNAG_04807 3 flanking region primer 2 <400> 1142 ggatttccga agccaaac 18 <210> 1143 <211> 19 <212> DNA <213> CNAG_04807 diagnostic screening primer, pairing with B79 <400> 1143 tggcacagtt caatgttcc 19 <210> 1144 <211> 18 <212> DNA <213> CNAG_04807 Southern blot probe primer <400> 1144 cgacaataca aacacgcc 18 <210> 1145 <211> 20 <212> DNA <213> NAT#177 STM primer <400> 1145 caccaactcc ccatctccat 20 <210> 1146 <211> 24 <212> DNA <213> STM common primer <400> 1146 gcatgccctg cccctaagaa ttcg 24 <210> 1147 <211> 19 <212> DNA <213> CNAG_02435 5 flanking region primer 1 <400> 1147 atcgttagca gggtgagtc 19 <210> 1148 <211> 42 <212> DNA <213> CNAG_02435 5 flanking region primer 2 <400> 1148 gctcactggc cgtcgtttta ctcctattca gggactatca cc 42 <210> 1149 <211> 39 <212> DNA <213> CNAG_02435 3 flanking region primer 1 <400> 1149 catggtcata gctgtttcct gacctatttt ccacgcagc 39 <210> 1150 <211> 18 <212> DNA <213> CNAG_02435 3 flanking region primer 2 <400> 1150 tcagcctcgc acataatc 18 <210> 1151 <211> 18 <212> DNA <213> CNAG_02435 diagnostic screening primer, pairing with B79 <400> 1151 cgagccgaaa agaacaag 18 <210> 1152 <211> 20 <212> DNA <213> CNAG_02435 Southern blot probe primer <400> 1152 gattatctgt ttggttcggc 20 <210> 1153 <211> 24 <212> DNA <213> NAT#184 STM primer <400> 1153 atatatggct cgagctagat agag 24 <210> 1154 <211> 24 <212> DNA <213> STM common primer <400> 1154 gcatgccctg cccctaagaa ttcg 24 <210> 1155 <211> 20 <212> DNA <213> CNAG_02364 5 flanking region primer 1 <400> 1155 gaagtcaaca ggaagaagcc 20 <210> 1156 <211> 34 <212> DNA <213> CNAG_02364 5 flanking region primer 2 <400> 1156 ctggccgtcg ttttactttg cgtgaaggtg tagg 34 <210> 1157 <211> 36 <212> DNA <213> CNAG_02364 3 flanking region primer 1 <400> 1157 gtcatagctg tttcctgtgc tcatcttcac ggctac 36 <210> 1158 <211> 19 <212> DNA <213> CNAG_02364 3 flanking region primer 2 <400> 1158 cattatccgc ctgatgttg 19 <210> 1159 <211> 19 <212> DNA <213> CNAG_02364 diagnostic screening primer, pairing with B79 <400> 1159 gcacttccaa caaagccac 19 <210> 1160 <211> 19 <212> DNA <213> CNAG_02364 Southern blot probe primer <400> 1160 gactttttgc gtgaaggtg 19 <210> 1161 <211> 24 <212> DNA <213> NAT#184 STM primer <400> 1161 atatatggct cgagctagat agag 24 <210> 1162 <211> 24 <212> DNA <213> STM common primer <400> 1162 gcatgccctg cccctaagaa ttcg 24 <210> 1163 <211> 20 <212> DNA <213> CNAG_05420 5 flanking region primer 1 <400> 1163 cgcctttgag gtattcactc 20 <210> 1164 <211> 39 <212> DNA <213> CNAG_05420 5 flanking region primer 2 <400> 1164 tcactggccg tcgttttacg gtgttggtca cgaatctac 39 <210> 1165 <211> 40 <212> DNA <213> CNAG_05420 3 flanking region primer 1 <400> 1165 catggtcata gctgtttcct gcggctttac gaaatgagtg 40 <210> 1166 <211> 21 <212> DNA <213> CNAG_05420 3 flanking region primer 2 <400> 1166 cctgaaagac atagcgacaa g 21 <210> 1167 <211> 19 <212> DNA <213> CNAG_05420 diagnostic screening primer, pairing with B79 <400> 1167 atcctgtctt gctgtggtc 19 <210> 1168 <211> 18 <212> DNA <213> CNAG_05420 Southern blot probe primer <400> 1168 tccagcatcg tcattgag 18 <210> 1169 <211> 20 <212> DNA <213> NAT#191 STM primer <400> 1169 atatggatgt ttttagcgag 20 <210> 1170 <211> 24 <212> DNA <213> STM common primer <400> 1170 gcatgccctg cccctaagaa ttcg 24 <210> 1171 <211> 19 <212> DNA <213> CNAG_05093 5 flanking region primer 1 <400> 1171 aatgtattgc caccgtgcc 19 <210> 1172 <211> 41 <212> DNA <213> CNAG_05093 5 flanking region primer 2 <400> 1172 tcactggccg tcgttttact tggggtgatg tgttctcgtc c 41 <210> 1173 <211> 41 <212> DNA <213> CNAG_05093 3 flanking region primer 1 <400> 1173 catggtcata gctgtttcct gacaggcaga ttgggaagtg c 41 <210> 1174 <211> 19 <212> DNA <213> CNAG_05093 3 flanking region primer 2 <400> 1174 tgtatgccgt gggaacaag 19 <210> 1175 <211> 24 <212> DNA <213> CNAG_05093 diagnostic screening primer, pairing with B79 <400> 1175 cgaagtctca aacctatcta ctcg 24 <210> 1176 <211> 22 <212> DNA <213> CNAG_05093 Southern blot probe primer <400> 1176 ttgctgaggc gaacttgttg gc 22 <210> 1177 <211> 24 <212> DNA <213> NAT#201 STM primer <400> 1177 caccctctat ctcgagaaag ctcc 24 <210> 1178 <211> 24 <212> DNA <213> STM common primer <400> 1178 gcatgccctg cccctaagaa ttcg 24 <210> 1179 <211> 19 <212> DNA <213> CNAG_06814 5 flanking region primer 1 <400> 1179 tgggaatggc aaaaggcac 19 <210> 1180 <211> 35 <212> DNA <213> CNAG_06814 5 flanking region primer 2 <400> 1180 ctggccgtcg ttttacggta ttttcgtcca gtaaa 35 <210> 1181 <211> 35 <212> DNA <213> CNAG_06814 3 flanking region primer 1 <400> 1181 gtcatagctg tttcctgtgc agcctatgcc tcctt 35 <210> 1182 <211> 19 <212> DNA <213> CNAG_06814 3 flanking region primer 2 <400> 1182 taagtgtgtt cacggtgcg 19 <210> 1183 <211> 19 <212> DNA <213> CNAG_06814 diagnostic screening primer, pairing with B79 <400> 1183 tggttaccga agtagggag 19 <210> 1184 <211> 19 <212> DNA <213> CNAG_06814 Southern blot probe primer <400> 1184 atttggaggt ggagttctg 19 <210> 1185 <211> 22 <212> DNA <213> NAT#208 STM primer <400> 1185 tggtcgcggg agatcgtggt tt 22 <210> 1186 <211> 24 <212> DNA <213> STM common primer <400> 1186 gcatgccctg cccctaagaa ttcg 24 <210> 1187 <211> 21 <212> DNA <213> CNAG_05431 5 flanking region primer 1 <400> 1187 catcagtctt gcttcttctg c 21 <210> 1188 <211> 40 <212> DNA <213> CNAG_05431 5 flanking region primer 2 <400> 1188 tcactggccg tcgttttaca gagtcagaat aggtgttggg 40 <210> 1189 <211> 40 <212> DNA <213> CNAG_05431 3 flanking region primer 1 <400> 1189 catggtcata gctgtttcct gagggagagg attgaagtgg 40 <210> 1190 <211> 20 <212> DNA <213> CNAG_05431 3 flanking region primer 2 <400> 1190 cattactttg actgggaggg 20 <210> 1191 <211> 19 <212> DNA <213> CNAG_05431 diagnostic screening primer, pairing with B79 <400> 1191 cacggcaact tatgctctc 19 <210> 1192 <211> 20 <212> DNA <213> CNAG_05431 Southern blot probe primer <400> 1192 actcggtgtt ggtgaaacgg 20 <210> 1193 <211> 22 <212> DNA <213> NAT#208 STM primer <400> 1193 tggtcgcggg agatcgtggt tt 22 <210> 1194 <211> 24 <212> DNA <213> STM common primer <400> 1194 gcatgccctg cccctaagaa ttcg 24 <210> 1195 <211> 20 <212> DNA <213> CNAG_05642 5 flanking region primer 1 <400> 1195 agagggataa gagcctgacg 20 <210> 1196 <211> 38 <212> DNA <213> CNAG_05642 5 flanking region primer 2 <400> 1196 tcactggccg tcgttttaca gaaggaagtt gggaagcg 38 <210> 1197 <211> 41 <212> DNA <213> CNAG_05642 3 flanking region primer 1 <400> 1197 catggtcata gctgtttcct gatgattaca gaggggagga c 41 <210> 1198 <211> 19 <212> DNA <213> CNAG_05642 3 flanking region primer 2 <400> 1198 tgtatcccaa gcagcaaag 19 <210> 1199 <211> 20 <212> DNA <213> CNAG_05642 diagnostic screening primer, pairing with B79 <400> 1199 ctcggaaata aatgtggagc 20 <210> 1200 <211> 20 <212> DNA <213> CNAG_05642 Southern blot probe primer <400> 1200 tacctgccag tgcttcaacc 20 <210> 1201 <211> 20 <212> DNA <213> NAT#210 STM primer <400> 1201 ctagagcccg ccacaacgct 20 <210> 1202 <211> 24 <212> DNA <213> STM common primer <400> 1202 gcatgccctg cccctaagaa ttcg 24 <210> 1203 <211> 19 <212> DNA <213> CNAG_03212 5 flanking region primer 1 <400> 1203 gacacccaaa caagcaaac 19 <210> 1204 <211> 39 <212> DNA <213> CNAG_03212 5 flanking region primer 2 <400> 1204 gctcactggc cgtcgtttta cctggggtgt ttgttttcg 39 <210> 1205 <211> 41 <212> DNA <213> CNAG_03212 3 flanking region primer 1 <400> 1205 catggtcata gctgtttcct ggggttcaga ttttggatac c 41 <210> 1206 <211> 19 <212> DNA <213> CNAG_03212 3 flanking region primer 2 <400> 1206 ggcgaggtgt tgtaaatcc 19 <210> 1207 <211> 22 <212> DNA <213> CNAG_03212 diagnostic screening primer, pairing with B79 <400> 1207 gcgtatctgg catcttgtta tc 22 <210> 1208 <211> 18 <212> DNA <213> CNAG_03212 Southern blot probe primer <400> 1208 tgaggaatgc caatggtg 18 <210> 1209 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 1209 gcggtcgctt tatagcgatt 20 <210> 1210 <211> 24 <212> DNA <213> STM common primer <400> 1210 gcatgccctg cccctaagaa ttcg 24 <210> 1211 <211> 18 <212> DNA <213> CNAG_03212 5 flanking region primer 1 <400> 1211 gcgaaacttt gtgagtgc 18 <210> 1212 <211> 40 <212> DNA <213> CNAG_03212 5 flanking region primer 2 <400> 1212 gctcactggc cgtcgtttta ccaagcagca agatggagtc 40 <210> 1213 <211> 40 <212> DNA <213> CNAG_03212 3 flanking region primer 1 <400> 1213 catggtcata gctgtttcct gtcgtaagtt cggtgtcagc 40 <210> 1214 <211> 20 <212> DNA <213> CNAG_03212 3 flanking region primer 2 <400> 1214 aagtcttatt atgggcgagg 20 <210> 1215 <211> 19 <212> DNA <213> CNAG_03212 diagnostic screening primer, pairing with B79 <400> 1215 tgtcccgttg ataggtgtc 19 <210> 1216 <211> 20 <212> DNA <213> CNAG_03212 Southern blot probe primer <400> 1216 tcctctccct cctattcagg 20 <210> 1217 <211> 20 <212> DNA <213> NAT#211 STM primer <400> 1217 gcggtcgctt tatagcgatt 20 <210> 1218 <211> 24 <212> DNA <213> STM common primer <400> 1218 gcatgccctg cccctaagaa ttcg 24 <210> 1219 <211> 19 <212> DNA <213> CNAG_07924 5 flanking region primer 1 <400> 1219 acctaatcag gaaagcccc 19 <210> 1220 <211> 39 <212> DNA <213> CNAG_07924 5 flanking region primer 2 <400> 1220 tcactggccg tcgttttacc attccttctc cgaaatctc 39 <210> 1221 <211> 40 <212> DNA <213> CNAG_07924 3 flanking region primer 1 <400> 1221 catggtcata gctgtttcct gttatcctcc cgaataccgc 40 <210> 1222 <211> 18 <212> DNA <213> CNAG_07924 3 flanking region primer 2 <400> 1222 ccgcaccaaa ctatttcg 18 <210> 1223 <211> 18 <212> DNA <213> CNAG_07924 diagnostic screening primer, pairing with B79 <400> 1223 atcgcacgaa aggcaaag 18 <210> 1224 <211> 20 <212> DNA <213> CNAG_07924 Southern blot probe primer <400> 1224 tctaaacccg tctctgactg 20 <210> 1225 <211> 20 <212> DNA <213> NAT#218 STM primer <400> 1225 ctccacatcc atcgctccaa 20 <210> 1226 <211> 24 <212> DNA <213> STM common primer <400> 1226 gcatgccctg cccctaagaa ttcg 24 <210> 1227 <211> 22 <212> DNA <213> CNAG_00156 5 flanking region primer 1 <400> 1227 cccgattttg ctttttagtg cg 22 <210> 1228 <211> 36 <212> DNA <213> CNAG_00156 5 flanking region primer 2 <400> 1228 ctggccgtcg ttttacttgc cgtctttgct tgggtc 36 <210> 1229 <211> 36 <212> DNA <213> CNAG_00156 3 flanking region primer 1 <400> 1229 gtcatagctg tttcctgatc aagtccactc ccgaag 36 <210> 1230 <211> 19 <212> DNA <213> CNAG_00156 3 flanking region primer <400> 1230 tagatgatga gggcgaacg 19 <210> 1231 <211> 20 <212> DNA <213> CNAG_00156 diagnostic screening primer, pairing with B79 <400> 1231 gcttcgttgg ttcgttagtc 20 <210> 1232 <211> 20 <212> DNA <213> CNAG_00156 Southern blot probe primer <400> 1232 caatccgttt ctctcacctc 20 <210> 1233 <211> 20 <212> DNA <213> NAT#231 STM primer <400> 1233 gagagatccc aacatcacgc 20 <210> 1234 <211> 24 <212> DNA <213> STM common primer <400> 1234 gcatgccctg cccctaagaa ttcg 24 <210> 1235 <211> 19 <212> DNA <213> CNAG_01173 5 flanking region primer 1 <400> 1235 agtatcccaa gcatccctc 19 <210> 1236 <211> 39 <212> DNA <213> CNAG_01173 5 flanking region primer 2 <400> 1236 tcactggccg tcgttttacg gcaggagcaa agtgtattc 39 <210> 1237 <211> 39 <212> DNA <213> CNAG_01173 3 flanking region primer 1 <400> 1237 catggtcata gctgtttcct gtatggtggc atctgaacc 39 <210> 1238 <211> 20 <212> DNA <213> CNAG_01173 3 flanking region primer 2 <400> 1238 ctgacaagtt taccaggacg 20 <210> 1239 <211> 20 <212> DNA <213> CNAG_01173 diagnostic screening primer, pairing with B79 <400> 1239 gcttctgtgt gtctcctctg 20 <210> 1240 <211> 20 <212> DNA <213> CNAG_01173 Southern blot probe primer <400> 1240 gagaaagaag gattccgaag 20 <210> 1241 <211> 24 <212> DNA <213> NAT#242 STM primer <400> 1241 gtagcgatag gggtgtcgct ttag 24 <210> 1242 <211> 24 <212> DNA <213> STM common primer <400> 1242 gcatgccctg cccctaagaa ttcg 24 <210> 1243 <211> 19 <212> DNA <213> CNAG_06871 5 flanking region primer 1 <400> 1243 tcaggatggg agtgtttgc 19 <210> 1244 <211> 37 <212> DNA <213> CNAG_06871 5 flanking region primer 2 <400> 1244 tcactggccg tcgttttacc cttctccttt tgcttgg 37 <210> 1245 <211> 41 <212> DNA <213> CNAG_06871 3 flanking region primer 1 <400> 1245 catggtcata gctgtttcct gcgctcttat taccgcttca g 41 <210> 1246 <211> 20 <212> DNA <213> CNAG_06871 3 flanking region primer 2 <400> 1246 tcggtatttc agtagggatg 20 <210> 1247 <211> 19 <212> DNA <213> CNAG_06871 diagnostic screening primer, pairing with B79 <400> 1247 cgactgctgg atgtatgag 19 <210> 1248 <211> 18 <212> DNA <213> CNAG_06871 Southern blot probe primer <400> 1248 cttgactgcc tcatctgc 18 <210> 1249 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 1249 acacctacat caaaccctcc c 21 <210> 1250 <211> 24 <212> DNA <213> STM common primer <400> 1250 gcatgccctg cccctaagaa ttcg 24 <210> 1251 <211> 18 <212> DNA <213> CNAG_04774 5 flanking region primer 1 <400> 1251 atttgaatag ccgcaccc 18 <210> 1252 <211> 35 <212> DNA <213> CNAG_04774 5 flanking region primer 2 <400> 1252 ctggccgtcg ttttacattg ggggatgctg gaaag 35 <210> 1253 <211> 36 <212> DNA <213> CNAG_04774 3 flanking region primer 1 <400> 1253 gtcatagctg tttcctgggt ggactgccaa actatg 36 <210> 1254 <211> 19 <212> DNA <213> CNAG_04774 3 flanking region primer 2 <400> 1254 aggcatttct gttcctccg 19 <210> 1255 <211> 19 <212> DNA <213> CNAG_04774 diagnostic screening primer, pairing with B79 <400> 1255 cgatagacgg aggatgaag 19 <210> 1256 <211> 18 <212> DNA <213> CNAG_04774 Southern blot probe primer <400> 1256 atcgttgctt ccgcttag 18 <210> 1257 <211> 21 <212> DNA <213> NAT#146 STM primer <400> 1257 actagccccc cctcaccacc t 21 <210> 1258 <211> 24 <212> DNA <213> STM common primer <400> 1258 gcatgccctg cccctaagaa ttcg 24 <210> 1259 <211> 18 <212> DNA <213> CNAG_00841 5 flanking region primer 1 <400> 1259 tcggcaagtt tagcaagc 18 <210> 1260 <211> 39 <212> DNA <213> CNAG_00841 5 flanking region primer 2 <400> 1260 gctcactggc cgtcgtttta cttatttctg ggctgtcgc 39 <210> 1261 <211> 40 <212> DNA <213> CNAG_00841 3 flanking region primer 1 <400> 1261 catggtcata gctgtttcct gtctgtggag tcagccaaac 40 <210> 1262 <211> 23 <212> DNA <213> CNAG_00841 3 flanking region primer 2 <400> 1262 cggtatcctc gtgagtctat ttc 23 <210> 1263 <211> 21 <212> DNA <213> CNAG_00841 diagnostic screening primer, pairing with B79 <400> 1263 gcagttgctg aagtgaaaga c 21 <210> 1264 <211> 20 <212> DNA <213> CNAG_00841 Southern blot probe primer <400> 1264 tgggatagaa gaaggctgtg 20 <210> 1265 <211> 24 <212> DNA <213> NAT#205 STM primer <400> 1265 tatccccctc tccgctctct agca 24 <210> 1266 <211> 24 <212> DNA <213> STM common primer <400> 1266 gcatgccctg cccctaagaa ttcg 24 <210> 1267 <211> 18 <212> DNA <213> CNAG_04836 5 flanking region primer 1 <400> 1267 tagaaaagcc acaacccc 18 <210> 1268 <211> 37 <212> DNA <213> CNAG_04836 5 flanking region primer 2 <400> 1268 tcactggccg tcgttttacg agtttctttt ggggtgg 37 <210> 1269 <211> 40 <212> DNA <213> CNAG_04836 3 flanking region primer 1 <400> 1269 catggtcata gctgtttcct gacgagaagg ggaatgatgg 40 <210> 1270 <211> 19 <212> DNA <213> CNAG_04836 3 flanking region primer 2 <400> 1270 tgggcatctt gctcattac 19 <210> 1271 <211> 19 <212> DNA <213> CNAG_04836 diagnostic screening primer, pairing with B79 <400> 1271 cggagagcaa atagcacag 19 <210> 1272 <211> 20 <212> DNA <213> CNAG_04836 Southern blot probe primer <400> 1272 aggtgtcgta tcgtatttgc 20 <210> 1273 <211> 20 <212> DNA <213> NAT#123 STM primer <400> 1273 ctatcgacca accaacacag 20 <210> 1274 <211> 24 <212> DNA <213> STM common primer <400> 1274 gcatgccctg cccctaagaa ttcg 24 <210> 1275 <211> 19 <212> DNA <213> CNAG_02603 5 flanking region primer 1 <400> 1275 tcaaaacgcc tttcggaac 19 <210> 1276 <211> 40 <212> DNA <213> CNAG_02603 5 flanking region primer 2 <400> 1276 gctcactggc cgtcgtttta cacaaatggc agttccctcg 40 <210> 1277 <211> 43 <212> DNA <213> CNAG_02603 3 flanking region primer 1 <400> 1277 catggtcata gctgtttcct gatctctccc tcacttcagt cgg 43 <210> 1278 <211> 19 <212> DNA <213> CNAG_02603 3 flanking region primer 2 <400> 1278 taggaccaac ttcagaccg 19 <210> 1279 <211> 20 <212> DNA <213> CNAG_02603 diagnostic screening primer, pairing with B79 <400> 1279 gcctatggaa gaagacaacg 20 <210> 1280 <211> 20 <212> DNA <213> CNAG_02603 Southern blot probe primer <400> 1280 gtcgtgtagt tttgaatggc 20 <210> 1281 <211> 19 <212> DNA <213> NAT#224 STM primer <400> 1281 aacctttaaa tgggtagag 19 <210> 1282 <211> 24 <212> DNA <213> STM common primer <400> 1282 gcatgccctg cccctaagaa ttcg 24 <210> 1283 <211> 20 <212> DNA <213> CNAG_03059 5 flanking region primer 1 <400> 1283 aatggcgtta ccgacgagag 20 <210> 1284 <211> 36 <212> DNA <213> CNAG_03059 5 flanking region primer 2 <400> 1284 ctggccgtcg ttttacggta ttgagagtgc cgagtg 36 <210> 1285 <211> 38 <212> DNA <213> CNAG_03059 3 flanking region primer 1 <400> 1285 gtcatagctg tttcctgcag actccctctt cttacctg 38 <210> 1286 <211> 24 <212> DNA <213> CNAG_03059 3 flanking region primer 2 <400> 1286 cgttactgcg aggttattac tgtg 24 <210> 1287 <211> 19 <212> DNA <213> CNAG_03059 diagnostic screening primer, pairing with B79 <400> 1287 aaagactcac ccttcaacg 19 <210> 1288 <211> 20 <212> DNA <213> CNAG_03059 Southern blot probe primer <400> 1288 cttttttccg ctggtttctc 20 <210> 1289 <211> 18 <212> DNA <213> NAT#232 STM primer <400> 1289 ctttaaaggt ggtttgtg 18 <210> 1290 <211> 24 <212> DNA <213> STM common primer <400> 1290 gcatgccctg cccctaagaa ttcg 24 <210> 1291 <211> 21 <212> DNA <213> CNAG_00883 5 flanking region primer 1 <400> 1291 cgacagaaac aagtccaagt g 21 <210> 1292 <211> 36 <212> DNA <213> CNAG_00883 5 flanking region primer 2 <400> 1292 ctggccgtcg ttttacgtag tgaaatgagg attgcg 36 <210> 1293 <211> 35 <212> DNA <213> CNAG_00883 3 flanking region primer 1 <400> 1293 gtcatagctg tttcctgtac ctgcttttct tgcgg 35 <210> 1294 <211> 18 <212> DNA <213> CNAG_00883 3 flanking region primer 2 <400> 1294 acaagcccga atctcaag 18 <210> 1295 <211> 19 <212> DNA <213> CNAG_00883 diagnostic screening primer, pairing with B79 <400> 1295 cttttcactc aatgtcccg 19 <210> 1296 <211> 19 <212> DNA <213> CNAG_00883 Southern blot probe primer <400> 1296 taagaactgt tgccctgcc 19 <210> 1297 <211> 18 <212> DNA <213> CNAG_01551 5 flanking region primer 1 <400> 1297 tccgttgaga tagcgttg 18 <210> 1298 <211> 39 <212> DNA <213> CNAG_01551 5 flanking region primer 2 <400> 1298 tcactggccg tcgttttaca tggtggaggt gtaggactg 39 <210> 1299 <211> 39 <212> DNA <213> CNAG_01551 3 flanking region primer 1 <400> 1299 catggtcata gctgtttcct gttattcccc ctcaagagc 39 <210> 1300 <211> 20 <212> DNA <213> CNAG_01551 3 flanking region primer 2 <400> 1300 gcggcccatt cttcttgtcg 20 <210> 1301 <211> 18 <212> DNA <213> CNAG_01551 diagnostic screening primer, pairing with B79 <400> 1301 aaccttgcct aacaaccc 18 <210> 1302 <211> 20 <212> DNA <213> CNAG_01551 Southern blot probe primer <400> 1302 ggagtatggc tgaaatctgg 20 <210> 1303 <211> 24 <212> DNA <213> NAT#273 STM primer <400> 1303 gagatctttc gggaggtctg gatt 24 <210> 1304 <211> 24 <212> DNA <213> STM common primer <400> 1304 gcatgccctg cccctaagaa ttcg 24 <210> 1305 <211> 20 <212> DNA <213> CNAG_04353 5 flanking region primer 1 <400> 1305 tgcgagagag aatgggagac 20 <210> 1306 <211> 38 <212> DNA <213> CNAG_04353 5 flanking region primer 2 <400> 1306 tcactggccg tcgttttacg gttccatagc gacatcag 38 <210> 1307 <211> 41 <212> DNA <213> CNAG_04353 3 flanking region primer 1 <400> 1307 catggtcata gctgtttcct gcgaaggttg tttgtgatac g 41 <210> 1308 <211> 20 <212> DNA <213> CNAG_04353 3 flanking region primer 2 <400> 1308 cccaacaact catcaatctc 20 <210> 1309 <211> 18 <212> DNA <213> CNAG_04353 diagnostic screening primer, pairing with B79 <400> 1309 gcattatcgt ttcccgac 18 <210> 1310 <211> 22 <212> DNA <213> CNAG_04353 Southern blot probe primer <400> 1310 ttgaggtagg tggagtgttg cc 22 <210> 1311 <211> 20 <212> DNA <213> CNAG_03346 5 flanking region primer 1 <400> 1311 cgccctttct attgttacac 20 <210> 1312 <211> 39 <212> DNA <213> CNAG_03346 5 flanking region primer 2 <400> 1312 tcactggccg tcgttttacg atgacaggag ggatgaatc 39 <210> 1313 <211> 43 <212> DNA <213> CNAG_03346 3 flanking region primer 1 <400> 1313 catggtcata gctgtttcct ggggagaata acgactcaat gtc 43 <210> 1314 <211> 18 <212> DNA <213> CNAG_03346 3 flanking region primer 2 <400> 1314 tcattgctga ctgggaag 18 <210> 1315 <211> 18 <212> DNA <213> CNAG_03346 diagnostic screening primer, pairing with B79 <400> 1315 aaagaggcgg tgttgaag 18 <210> 1316 <211> 19 <212> DNA <213> CNAG_03346 Southern blot probe primer <400> 1316 agccaggtaa tcttggagg 19 <210> 1317 <211> 21 <212> DNA <213> NAT#295 STM primer <400> 1317 acacctacat caaaccctcc c 21 <210> 1318 <211> 24 <212> DNA <213> STM common primer <400> 1318 gcatgccctg cccctaagaa ttcg 24 <210> 1319 <211> 18 <212> DNA <213> CNAG_00239 5 flanking region primer 1 <400> 1319 aggattatcg cctgaacc 18 <210> 1320 <211> 37 <212> DNA <213> CNAG_00239 5 flanking region primer 2 <400> 1320 tcactggccg tcgttttaca tgttgttgaa gagggcg 37 <210> 1321 <211> 41 <212> DNA <213> CNAG_00239 3 flanking region primer 1 <400> 1321 catggtcata gctgtttcct gaacgacgac catcgcagta g 41 <210> 1322 <211> 18 <212> DNA <213> CNAG_00239 3 flanking region primer 2 <400> 1322 catcccgagt tggaaaac 18 <210> 1323 <211> 20 <212> DNA <213> CNAG_00239 diagnostic screening primer, pairing with B79 <400> 1323 cactggcgaa gtcaagtatg 20 <210> 1324 <211> 20 <212> DNA <213> CNAG_00239 Southern blot probe primer <400> 1324 ctactctgtg gtggcgtaag 20 <210> 1325 <211> 20 <212> DNA <213> NAT#296 STM primer <400> 1325 cgcccgccct cactatccac 20 <210> 1326 <211> 24 <212> DNA <213> STM common primer <400> 1326 gcatgccctg cccctaagaa ttcg 24 <210> 1327 <211> 19 <212> DNA <213> CNAG_03129 5 flanking region primer 1 <400> 1327 gcaggcgaaa actattagg 19 <210> 1328 <211> 41 <212> DNA <213> CNAG_03129 5 flanking region primer 2 <400> 1328 gctcactggc cgtcgtttta cgtggggaag tcctcaaata c 41 <210> 1329 <211> 42 <212> DNA <213> CNAG_03129 3 flanking region primer 1 <400> 1329 catggtcata gctgtttcct gcgacgacat ctcaagtcag ag 42 <210> 1330 <211> 20 <212> DNA <213> CNAG_03129 3 flanking region primer 2 <400> 1330 tgatgggacg ataaaggttc 20 <210> 1331 <211> 19 <212> DNA <213> CNAG_03129 diagnostic screening primer, pairing with B79 <400> 1331 tgctttctct aatagcggc 19 <210> 1332 <211> 19 <212> DNA <213> CNAG_03129 Southern blot probe primer <400> 1332 gtgcttttga agttccgtc 19 <210> 1333 <211> 18 <212> DNA <213> CNAG_03826 5 flanking region primer 1 <400> 1333 aatgctgatt tggcaggg 18 <210> 1334 <211> 39 <212> DNA <213> CNAG_03826 5 flanking region primer 2 <400> 1334 tcactggccg tcgttttacc gatggaagat gttgtcagc 39 <210> 1335 <211> 39 <212> DNA <213> CNAG_03826 3 flanking region primer 1 <400> 1335 catggtcata gctgtttcct gacaaagcga aaggcagac 39 <210> 1336 <211> 18 <212> DNA <213> CNAG_03826 3 flanking region primer 2 <400> 1336 cgaaagtgat gcttcagc 18 <210> 1337 <211> 20 <212> DNA <213> CNAG_03826 diagnostic screening primer, pairing with B79 <400> 1337 ggaaggtcag gtcctatttg 20 <210> 1338 <211> 20 <212> DNA <213> CNAG_02671 5 flanking region primer 1 <400> 1338 cgccgagtta tctgacaaag 20 <210> 1339 <211> 41 <212> DNA <213> CNAG_02671 5 flanking region primer 2 <400> 1339 tcactggccg tcgttttact tctccaaaca ccgcccttga c 41 <210> 1340 <211> 40 <212> DNA <213> CNAG_02671 3 flanking region primer 1 <400> 1340 catggtcata gctgtttcct gtaacggggc ttagaagtgc 40 <210> 1341 <211> 18 <212> DNA <213> CNAG_02671 3 flanking region primer 2 <400> 1341 ttcatcaaag gtctggcg 18 <210> 1342 <211> 20 <212> DNA <213> CNAG_02671 diagnostic screening primer, pairing with B79 <400> 1342 cctgactcat cggcttactc 20 <210> 1343 <211> 18 <212> DNA <213> CNAG_02671 Southern blot probe primer <400> 1343 actcctgctt gcgttttc 18 <210> 1344 <211> 20 <212> DNA <213> CNAG_02134 5 flanking region primer 1 <400> 1344 ggggtaacta tcatcatccg 20 <210> 1345 <211> 38 <212> DNA <213> CNAG_02134 5 flanking region primer 2 <400> 1345 tcactggccg tcgttttact ctcgctcctt ttccttcc 38 <210> 1346 <211> 42 <212> DNA <213> CNAG_02134 3 flanking region primer 1 <400> 1346 catggtcata gctgtttcct ggcagtagaa ggtgatacaa cg 42 <210> 1347 <211> 19 <212> DNA <213> CNAG_02134 3 flanking region primer 2 <400> 1347 cgttacggtt ggacaatgg 19 <210> 1348 <211> 19 <212> DNA <213> CNAG_02134 diagnostic screening primer, pairing with B79 <400> 1348 cgtagaaaca acgcacgag 19 <210> 1349 <211> 21 <212> DNA <213> CNAG_02134 Southern blot probe primer <400> 1349 caagtattct ccagggttca g 21 <210> 1350 <211> 20 <212> DNA <213> CNAG_02936 5 flanking region primer 1 <400> 1350 gcgagtgatg acgagagttc 20 <210> 1351 <211> 39 <212> DNA <213> CNAG_02936 5 flanking region primer 2 <400> 1351 tcactggccg tcgttttacg ccttgaccgt ctcttctac 39 <210> 1352 <211> 42 <212> DNA <213> CNAG_02936 3 flanking region primer 1 <400> 1352 catggtcata gctgtttcct ggtctactcg tcactttcat cg 42 <210> 1353 <211> 21 <212> DNA <213> CNAG_02936 3 flanking region primer 2 <400> 1353 cgtcacaaca agcactctca c 21 <210> 1354 <211> 19 <212> DNA <213> CNAG_02936 diagnostic screening primer, pairing with B79 <400> 1354 ggatgttcag gtgccaatc 19 <210> 1355 <211> 18 <212> DNA <213> CNAG_07460 5 flanking region primer 1 <400> 1355 tcggcattgg catacttg 18 <210> 1356 <211> 39 <212> DNA <213> CNAG_07460 5 flanking region primer 2 <400> 1356 gctcactggc cgtcgtttta cgtgatttga gcggtgaag 39 <210> 1357 <211> 42 <212> DNA <213> CNAG_07460 3 flanking region primer 1 <400> 1357 catggtcata gctgtttcct gccaaagtgg actcaaatag gg 42 <210> 1358 <211> 22 <212> DNA <213> CNAG_07460 3 flanking region primer 2 <400> 1358 ccgttacttt ccaaatcagg ag 22 <210> 1359 <211> 20 <212> DNA <213> CNAG_07460 diagnostic screening primer, pairing with B79 <400> 1359 cgtctgaacc tgattctgag 20 <210> 1360 <211> 19 <212> DNA <213> CNAG_07460 Southern blot probe primer <400> 1360 ccgatgacca aaagtaggc 19 <210> 1361 <211> 19 <212> DNA <213> CNAG_07460 Southern blot probe primer <400> 1361 cagagttgat tgccgaaag 19 <210> 1362 <211> 19 <212> DNA <213> CNAG_05436 5 flanking region primer 1 <400> 1362 gcccttttgc tttaccttc 19 <210> 1363 <211> 41 <212> DNA <213> CNAG_05436 5 flanking region primer 2 <400> 1363 tcactggccg tcgttttacc gtagagggat gattttgaga c 41 <210> 1364 <211> 40 <212> DNA <213> CNAG_05436 3 flanking region primer 1 <400> 1364 catggtcata gctgtttcct ggcatccttc gtgctaacag 40 <210> 1365 <211> 18 <212> DNA <213> CNAG_05436 3 flanking region primer 2 <400> 1365 gccttgaaga acaaacgg 18 <210> 1366 <211> 19 <212> DNA <213> CNAG_05436 diagnostic screening primer, pairing with B79 <400> 1366 gggataggag gcattgatg 19 <210> 1367 <211> 20 <212> DNA <213> CNAG_05436 Southern blot probe primer <400> 1367 caccaaagtg gatttccttg 20 <210> 1368 <211> 21 <212> DNA <213> CNAG_05436 Southern blot probe primer <400> 1368 ctcggacaaa tcactctact g 21 <210> 1369 <211> 21 <212> DNA <213> CNAG_02698 5 flanking region primer 1 <400> 1369 ctacagataa tgtgctcagc c 21 <210> 1370 <211> 37 <212> DNA <213> CNAG_02698 5 flanking region primer 2 <400> 1370 tcactggccg tcgttttact ttgttccata gccagcg 37 <210> 1371 <211> 40 <212> DNA <213> CNAG_02698 3 flanking region primer 1 <400> 1371 catggtcata gctgtttcct ggttctgtct tgaagggtgc 40 <210> 1372 <211> 20 <212> DNA <213> CNAG_02698 3 flanking region primer 2 <400> 1372 cggtgaagtt ggctaagaac 20 <210> 1373 <211> 19 <212> DNA <213> CNAG_02698 diagnostic screening primer, pairing with B79 <400> 1373 gaaagggcta agaagcgac 19 <210> 1374 <211> 20 <212> DNA <213> CNAG_02698 Southern blot probe primer <400> 1374 aaactctcca tctgtgtccc 20 <210> 1375 <211> 20 <212> DNA <213> CNAG_04600 5 flanking region primer 1 <400> 1375 gctgttggct gattgtttac 20 <210> 1376 <211> 39 <212> DNA <213> CNAG_04600 5 flanking region primer 2 <400> 1376 tcactggccg tcgttttact tacagtatcc caccgtccc 39 <210> 1377 <211> 40 <212> DNA <213> CNAG_04600 3 flanking region primer 1 <400> 1377 catggtcata gctgtttcct gtgggagtga tgtctttggc 40 <210> 1378 <211> 18 <212> DNA <213> CNAG_04600 3 flanking region primer 2 <400> 1378 ccctcaaaag ttgcgaag 18 <210> 1379 <211> 18 <212> DNA <213> CNAG_04600 diagnostic screening primer, pairing with B79 <400> 1379 ctggattttg gaatgggc 18 <210> 1380 <211> 18 <212> DNA <213> CNAG_04600 Southern blot probe primer <400> 1380 ttacatttgc cagcggac 18
Claims (21)
(b) 상기 세포에서 독성 조절 유전자의 발현을 측정하는 단계;
(c) 상기 단계에서 독성 조절 유전자의 발현이 감소조절 (down-regulation)되는 것으로 측정될 때, 상기 시료가 항균제임을 판별하는 단계를 포함하는 항균제 스크리닝 방법. (a) a toxicity control genes that regulate the pathogenic cellular usv101, fzc1, hob1, zfc2, fzc50, fzc31, bzp2, fzc9, ddt1, mal13, fzc2, fzc43, fzc22, hlh1, mbs2, rum1, fzc5, aro80, clr1 , pip2 , fzc37 , gat5 , fzc49 , cef3 , fzc33 , fzc12, and zfc5 to a cell of a cryptococcus neoformans cell containing the gene to be analyzed;
(b) measuring the expression of a toxic regulatory gene in the cell;
(c) determining that the sample is an antimicrobial agent when the expression of the toxic regulatory gene is determined to be down-regulated in the step (c).
(b) 상기 세포에서 독성 조절 유전자의 발현을 측정하는 단계;
(c) 상기 단계에서 독성 조절 유전자의 발현이 증가조절 (up-regulation)되는 것으로 측정될 때, 상기 시료가 항균제임을 판별하는 단계를 포함하는 항균제 스크리닝 방법. (a) a toxic regulatory gene that regulates the virulence of a cell, including a gene selected from the group consisting of fzc17 , fzc40 , aro8001 , fzc38 , fzc24, and ertl ; cryptococcus contacting neoformans cells with a sample to be analyzed;
(b) measuring the expression of a toxic regulatory gene in the cell;
(c) determining that the sample is an antimicrobial agent when the expression of the toxic regulatory gene is measured to be up-regulated in the step (a).
상기 멜라닌 생산을 조절하는 유전자는 bzp2, fkh2, bzp3, hlh1, sip4, rds2, sip401, fzc1, gat1, ada2, nrg1, fzc31 및 hlh2로 구성된 군으로부터 선택된 어느 하나의 유전자이며,
상기 우레아제 생산을 조절하는 유전자는 rim1, atf1, fkh2, fzc1, usv101, sxi1alpha, mln1, fzc26, skn7, zfc7, hob7, fzc14 및 hob4로 구성된 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 항균제 스크리닝 방법.7. The method according to claim 5, wherein the gene for controlling the production of the capsules is selected from the group consisting of hob7 , clr3 , fzc51 , fzc1 , fkh2 , nrg1 , usv101 , fzc29 , bzp3 , zfc3 , fzc14 , sre1 , fzc30 , hlh4 , fzc36 , cr16 , mln1 , fzc46 , clr1 , fzc17 , jjj1 , fzc49 , fzc18 , hcm1 , fzc24 , hlh3 and hpa1 ,
Wherein the gene for controlling melanin production is any gene selected from the group consisting of bzp2 , fkh2 , bzp3 , hlhl , sip4 , rds2 , sip401 , fzc1 , gat1 , ada2 , nrg1 , fzc31 and hlh2 ,
Wherein the gene regulating the production of urease is any gene selected from the group consisting of rim1 , atf1 , fkh2 , fzc1 , usv101 , sxi1alpha , mln1 , fzc26 , skn7 , zfc7 , hob7 , fzc14 and hob4 .
(b) 상기 세포에서 독성 조절 유전자의 발현을 측정하는 단계;
(c) 상기 단계에서 독성 조절 유전자의 발현이 감소조절 (down-regulation)되는 것으로 측정될 때, 상기 시료가 뇌수막염 치료제임을 판별하는 단계를 포함하는 뇌수막염 치료제 스크리닝 방법.(a) a toxicity control genes that regulate the pathogenic cellular usv101, fzc1, hob1, zfc2, fzc50, fzc31, bzp2, fzc9, ddt1, mal13, fzc2, fzc43, fzc22, hlh1, mbs2, rum1, fzc5, aro80, clr1 , pip2 , fzc37 , gat5 , fzc49 , cef3 , fzc33 , fzc12, and zfc5 to a cell of the cryptococcus neoformans cell;
(b) measuring the expression of a toxic regulatory gene in the cell;
(c) determining that the sample is a treatment for meningococcal disease when the expression of the toxic control gene is down-regulated in the step (c).
상기 멜라닌 생산을 조절하는 유전자는 bzp4, fzc8, hob1, usv101, liv1, mbs2, fzc5, fzc25 및 ert1로 구성된 군으로부터 선택된 어느 하나의 유전자이며,
상기 우레아제 생산을 조절하는 유전자는 zap104, sre1, gat201, fzc46, hlh1 및 fzc21로 구성된 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 뇌수막염 치료제 스크리닝 방법.10. The method of claim 9, wherein the gene that regulates the production of capsules is selected from the group consisting of rds2 , zap104 , fzc47 , gat204 , fzc33 , fzc45 , hsf2 , bzp4 , hob5 , fzc16 , hob3 , zfc4 , mcm1 , liv4 , hob4 and liv1 Any one gene,
The gene controlling melanin production is any gene selected from the group consisting of bzp4 , fzc8 , hob1 , usv101 , liv1 , mbs2 , fzc5 , fzc25 and ert1 ,
Wherein the gene that regulates urease production is any one selected from the group consisting of zap104 , sre1 , gat201 , fzc46 , hlh1, and fzc21 .
(b) 상기 세포에서 독성 조절 유전자의 발현을 측정하는 단계;
(c) 상기 단계에서 독성 조절 유전자의 발현이 증가조절 (up-regulation)되는 것으로 측정될 때, 상기 시료가 뇌수막염 치료제임을 판별하는 단계를 포함하는 뇌수막염 치료제 스크리닝 방법.(a) a toxic regulatory gene that regulates the virulence of a cell, including a gene selected from the group consisting of fzc17 , fzc40 , aro8001 , fzc38 , fzc24, and ertl ; cryptococcus contacting neoformans cells with a sample to be analyzed;
(b) measuring the expression of a toxic regulatory gene in the cell;
(c) determining that the sample is a therapeutic agent for meningococcal when the expression of the toxic regulatory gene is measured as up-regulation in the step (c).
상기 멜라닌 생산을 조절하는 유전자는 bzp2, fkh2, bzp3, hlh1, sip4, rds2, sip401, fzc1, gat1, ada2, nrg1, fzc31 및 hlh2로 구성된 군으로부터 선택된 어느 하나의 유전자이며,
상기 우레아제 생산을 조절하는 유전자는 rim1, atf1, fkh2, fzc1, usv101, sxi1alpha, mln1, fzc26, skn7, zfc7, hob7, fzc14 및 hob4로 구성된 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 뇌수막염 치료제 스크리닝 방법.13. The method according to claim 12, wherein the gene that regulates the production of the capsules is selected from the group consisting of hob7 , clr3 , fzc51 , fzc1 , fkh2 , nrg1 , usv101 , fzc29 , bzp3 , zfc3 , fzc14 , sre1 , fzc30 , hlh4 , fzc36 , cr16 , mln1 , clr1 , fzc17 , jjj1 , fzc49 , fzc18 , hcm1 , fzc24 , hlh3 and hpa1 ,
Wherein the gene for controlling melanin production is any gene selected from the group consisting of bzp2 , fkh2 , bzp3 , hlhl , sip4 , rds2 , sip401 , fzc1 , gat1 , ada2 , nrg1 , fzc31 and hlh2 ,
Wherein the gene regulating the production of urease is any one selected from the group consisting of rim1 , atf1 , fkh2 , fzc1 , usv101 , sxi1alpha , mln1 , fzc26 , skn7 , zfc7 , hob7 , fzc14 and hob4 . Way.
(a) 단계의 세포는 기탁번호 KCCM51291로 기탁된 세포주 라이브러리로부터 선택된 어느 하나의 세포인 것을 특징으로 하는 뇌수막염 치료제 스크리닝 방법.13. The method according to claim 11 or 12,
wherein the cell of step (a) is any one selected from a cell line library deposited with accession number KCCM51291.
(b) 상기 독성 조절 유전자를 포함하는 크립토코커스 네오포만스 (cryptococcus neoformans) 세포에 분석할 시료 및 상기 항균제를 접촉시키고, 상기 유전자의 발현량을 측정하는 제 2 측정 단계; 및
(c) 제 1 측정 단계 및 제 2 측정 단계의 측정값을 비교하여, 제 2 측정 단계의 측정값이 제1 측정 단계의 측정값보다 감소조절 (down-regulation)될 때, 상기 시료가 병용 투여용 항균제임을 판별하는 병용 투여용 항균제 스크리닝 방법.(a) a toxicity control genes that regulate the pathogenic cellular usv101, fzc1, hob1, zfc2, fzc50, fzc31, bzp2, fzc9, ddt1, mal13, fzc2, fzc43, fzc22, hlh1, mbs2, rum1, fzc5, aro80, clr1 a method of contacting an antimicrobial agent with cryptococcus neoformans cells comprising a gene selected from the group consisting of pip2 , fzc37 , gat5 , fzc49 , cef3 , fzc33 , fzc12 and zfc5 , 1 measuring step;
(b) a second measuring step of contacting a sample to be analyzed with cryptococcus neoformans cells containing the toxic control gene and the antimicrobial agent and measuring the expression level of the gene; And
(c) comparing the measured values of the first measurement step and the second measurement step, and when the measured value of the second measurement step is down-regulated more than the measured value of the first measurement step, Wherein the antimicrobial agent is an antimicrobial agent.
상기 멜라닌 생산을 조절하는 유전자는 bzp4, fzc8, hob1, usv101, liv1, mbs2, fzc5, fzc25 및 ert1로 구성된 군으로부터 선택된 어느 하나의 유전자이며,
상기 우레아제 생산을 조절하는 유전자는 zap104, sre1, gat201, fzc46, hlh1 및 fzc21로 구성된 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 병용 투여용 항균제 스크리닝 방법.22. The method of claim 21, wherein the gene that regulates the production of capsules is selected from the group consisting of rds2 , zap104 , fzc47 , gat204 , fzc33 , fzc45 , hsf2 , bzp4 , hob5 , fzc16 , hob3 , zfc4 , mcm1 , liv4 , hob4 and liv1 Any one gene,
The gene controlling melanin production is any gene selected from the group consisting of bzp4 , fzc8 , hob1 , usv101 , liv1 , mbs2 , fzc5 , fzc25 and ert1 ,
Wherein the gene regulating the production of urease is any gene selected from the group consisting of zap104 , sre1 , gat201 , fzc46 , hlh1 and fzc21 .
(b) 상기 독성 조절 유전자를 포함하는 크립토코커스 네오포만스 (cryptococcus neoformans) 세포에 분석할 시료 및 상기 항균제를 접촉시키고, 상기 유전자의 발현량을 측정하는 제 2 측정 단계; 및
(c) 제 1 측정 단계 및 제 2 측정 단계의 측정값을 비교하여, 제 2 측정 단계의 측정값이 제1 측정 단계의 측정값보다 증가조절 (up-regulation)될 때, 상기 시료가 병용 투여용 항균제임을 판별하는 병용 투여용 항균제 스크리닝 방법.(a) a toxic regulatory gene that regulates the virulence of a cell, including a gene selected from the group consisting of fzc17 , fzc40 , aro8001 , fzc38 , fzc24, and ertl ; cryptococcus a first measuring step of bringing an antimicrobial agent into contact with a neoformans cell and measuring an expression level of the gene;
(b) a second measuring step of contacting a sample to be analyzed with cryptococcus neoformans cells containing the toxic control gene and the antimicrobial agent and measuring the expression level of the gene; And
(c) comparing the measurement values of the first measurement step and the second measurement step, and when the measurement value of the second measurement step is up-regulated above the measurement value of the first measurement step, Wherein the antimicrobial agent is an antimicrobial agent.
상기 멜라닌 생산을 조절하는 유전자는 bzp2, fkh2, bzp3, hlh1, sip4, rds2, sip401, fzc1, gat1, ada2, nrg1, fzc31 및 hlh2로 구성된 군으로부터 선택된 어느 하나의 유전자이며,
상기 우레아제 생산을 조절하는 유전자는 rim1, atf1, fkh2, fzc1, usv101, sxi1alpha, mln1, fzc26, skn7, zfc7, hob7, fzc14 및 hob4로 구성된 군으로부터 선택된 어느 하나의 유전자인 것을 특징으로 하는 병용 투여용 항균제 스크리닝 방법.20. The method according to claim 19, wherein the gene regulating the production of the capsules is selected from the group consisting of hob7 , clr3 , fzc51 , fzc1 , fkh2 , nrg1 , usv101 , fzc29 , bzp3 , zfc3 , fzc14 , sre1 , fzc30 , hlh4 , fzc36 , cr16 , mln1 , fzc46 , clr1 , fzc17 , jjj1 , fzc49 , fzc18 , hcm1 , fzc24 , hlh3 and hpa1 ,
Wherein the gene for controlling melanin production is any gene selected from the group consisting of bzp2 , fkh2 , bzp3 , hlhl , sip4 , rds2 , sip401 , fzc1 , gat1 , ada2 , nrg1 , fzc31 and hlh2 ,
Wherein the gene regulating the production of urease is any gene selected from the group consisting of rim1 , atf1 , fkh2 , fzc1 , usv101 , sxi1alpha , mln1 , fzc26 , skn7 , zfc7 , hob7 , fzc14 and hob4 Method of screening antimicrobial agents.
(a) 단계 및 (b) 단계의 세포는 기탁번호 KCCM51291로 기탁된 세포주 라이브러리로부터 선택된 어느 하나의 세포인 것을 특징으로 하는 병용 투여용 항균제 스크리닝 방법.20. The method according to claim 18 or 19,
wherein the cells of step (a) and step (b) are cells selected from a cell line library deposited with accession number KCCM51291.
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