KR20170077066A - A target gene of hob1 for anti-fungal agent and the use thereof - Google Patents

Abstract

The present invention relates to antimicrobial target genes that regulate resistance to antimicrobial resistance, growth, crossing and external stress in Cryptococcus neoformans strains and uses thereof. According to the present invention, it is possible to effectively search candidate antimicrobial agents that detect the expression level of a novel transcription factor of the Cryptococcus neoformans strain and thereby increase the antibacterial effect or the therapeutic effect of meningitis. 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

[A target gene of hob1 for anti-fungal agent and the use thereof]

The present invention relates to target genes of Cryptococcus neoformans strains and uses thereof. In addition, the present invention relates to an antibacterial agent screening method, characterized in that the expression of the antimicrobial agent target gene of the Cryptococcus neoformans strain is confirmed.

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.

On the other hand, Cryptococcus 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 worldwide every year (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 Document 3, Non-Patent Document 4, and Non-Patent Document 5). The present inventors have verified that the Ire1 and Hxl1 (HAC1 and XBP1-like genes) proteins newly identified by Cryptococcus neoformans and the effect of antibacterial or meningitis treatment are effective in the case of impairing the function of the gene encoding them (Patent Document 1). , Patent Document 2). Most of the conventionally known signaling cascades are composed of sensor/receptor-like proteins and kinase/phosphatase, and contain unique adapter proteins or scaffold proteins to improve the specificity of each signaling pathway and to improve signaling pathways. Abnormal cross-talk between them was prevented. Nevertheless, each signaling cascade ultimately activates or inhibits a single transcription factor (TF) or multiple transcription factors, thereby regulating the level of transcription of the transcription factor. The effector protein of the transcription factor was increased-regulated or decreased-regulated by the binding of the transcription factor to. Thus, transcription factors are recognized as major regulators of gene expression within a given signaling pathway. In particular, the list of transcription factors is more granular than other signaling components between species. As described in the recent genome analysis results, in the case of Cryptococcus neoformans, transcription factors are further subdivided in particular (Non-Patent Document 6). For example, the UPR signaling pathway, which is important for endoplasmic reticulum (ER) stress response and toxicity of Cryptococcus neoformans, consists of an evolutionarily highly conserved Ire1 kinase and downstream Hxl1 transcription factor. Thus, Cryptococcus neoformans appears to have a number of evolutionarily conserved signaling cascades characterized by various types of transcription factors, which are unique Cryptococcus neoformans compared to other fungal signaling cascades. You can adjust the characteristics of the su.

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 Document 7 and Non-Patent Document 8). The prepared transcription factor knockout (TFKO) strains were analyzed for 30 different in vitro traits, which are related to growth, differentiation, stress responsiveness, antifungal resistance, and virulence factor production. Moreover, in the present invention, a large-scale toxicity test was performed using the digitization of Signature-Tag Mutagenesis (STM) in the insect host model and the murine host model, whereby various transcription factors in the Cryptococcus neoformans strain The present invention was completed by analyzing the trait according to the deletion and constructing a phenotypic dataset (prototype, phenome) for the transcription factor.

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.

Korean Registered Patent Publication No. 10-1311196 Korean Registered Patent Publication No. 10-1403862

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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.

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.

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.

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.

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.

In order to achieve the above object, the present inventors provide a Cryptococcus neoformans strain deposited under the accession number KCCM51291.

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 an antimicrobial agent target gene in Cryptococcus neoformans is measured.

In one embodiment, (a) contacting a sample to be analyzed with a cell line deposited with accession number KCCM51291 containing an antimicrobial target gene; (b) measuring the expression of an antimicrobial target gene in the cell line; (c) When it is determined that the expression of the antimicrobial target gene is down-regulated or up-regulated in the step, an antimicrobial agent screening method comprising the step of determining that the sample is an antimicrobial agent, wherein the The antimicrobial agent target gene provides an antimicrobial agent screening method, characterized in that it is any one gene selected from the group of genes consisting of an antimicrobial agent resistance regulating gene, a growth regulating gene, a mating regulating gene, and a response regulating gene for external stress.

In the above embodiment, the antimicrobial agent resistance regulating gene is a gene that regulates resistance to any one antimicrobial agent selected from the group consisting of azole-based, polyene-based, 5-flucytosine-based, and phenylpyrazole-based antimicrobial agents. It provides an antimicrobial agent screening method.

In the above embodiment, when the expression of the antimicrobial target gene is decreased, the genes that increase the sensitivity to azole-based antimicrobial agents are bzp3, hlh3, bzp/hxl1, sre1, rim101, bap2, hlh1, yap4, pip2, miz1, mln1, hob6. , mbf1, met32, fzc46, bap1, fzc14, fzc2, liv4, hsf2, zfc6, fzc45, fzc30, asg1, ste12, liv1, fzc22, fzc31, pan1, bzp2, sp1/crz1, bzp1 alpha, fzc34, hlh2, sxi Any one gene selected from the group consisting of fzc40, fzc38 and fzc17 , and genes that increase susceptibility to polyene-based antimicrobial agents are hob1, mbs1, jjj1, ert1, ecm22, gat201, zap104, sp1/crz1, fzc6, bzp5, hlh1, pip2, hcm1, bzp2, usv101, hob4, ste12, hob5, grf1, hel2, fzc45, asg1, fzc22, hob6, pan1, liv4, cuf1, fzc49, fzc1, bwc2, fap1, fzc44, fzc204, fzc204 , nrg1, pip201, hlh2, rim101, fzc38, hlh3, bzp3, mln1, met32, zfc2, fzc40, fzc31, and rum1 . The gene is any one selected from the group consisting of nrg1, zfc2, bap1, mbs1, fzc6, bap2, bzp3, jjj1, hlh1, pip2, apn2, fzc46, hap2, fzc51, bzp5, hcm1 and fzc19 , and the phenylpyrazole family Genes that increase susceptibility to antimicrobial agents are usv101, ada2, bap1, fzc6, hlh1, pip2, fzc46, hap2, bzp1/hxl1, fkh2, l iv1, bap2, bzp2, fzc21, hlh3, yrm101, bzp5, gln3, zfc8, ddt1, fzc22, hob6, rlm1, mln1, liv4, pan1, fzc35, yrm103, fzc3, asg1, fzc41, fzc51, hap1, fzc51, hap1, fzc41, fzc43, fzc22 It provides a method for screening an antimicrobial susceptibility enhancer, characterized in that it is any one gene selected from the group consisting of met32 and fzc32.

In the above embodiment, when the expression of the antimicrobial target gene is increased, genes that increase the sensitivity to azole-based antimicrobial agents are hob1, hap2, skn7, nrg1, mbs1, ppr1, jjj1, hcm1, ada2, fzc9, gat7, ert1, fkh2. , ecm22, ddt1, gat1, yrm103, cuf1 and fzc51 are any one gene selected from the group consisting of, and genes that increase susceptibility to polyene-based antimicrobial agents are sre1, bap1, fzc51, skn7, clr1, bzp5, atf1 and fzc4. It is any one gene selected from the group consisting of, and the gene that increases susceptibility to 5- flucytosine-based antimicrobial agents is any one gene selected from the group consisting of hlh3, rim101, gat204, hob3, fzc50, znf2, and rds2 , Genes that increase susceptibility to phenylpyrazole-based antimicrobial agents are any one gene selected from the group consisting of nrg1, jjj1, sp1/crz1, skn7, gat7, fap1, zfc2, gat204, znf2, hel2, fzc50, and sre1. It provides an antibacterial agent sensitivity enhancing agent screening screening method.

In the above embodiment, the growth regulation gene as the antimicrobial target gene by expression reduction is temperature independent or temperature dependent, and the temperature independent growth regulation gene is any one selected from the group consisting of bzp2, cuf1, hob1, gat5, fzc6 and nrg1. And the temperature-dependent growth regulation genes of 37°C to 39°C are hxl1, crz1, atf1, ada2, liv4, aro80, usv101, fzc31, fzc30, mln1, fzc30, fzc1, miz1, apn2, gat6, mbs2, sre1 and It provides an antibacterial agent screening method, characterized in that it is any one gene selected from the group consisting of ert1. In addition, it provides an antimicrobial agent screening method, characterized in that the antimicrobial agent target gene that regulates growth at 39°C as an antimicrobial agent target gene by increased expression is either a gene of mln1 or fzc46.

In the above embodiment, the mating control gene as the antimicrobial target gene by expression reduction is any one gene selected from the group consisting of bzp2, usv101, fzc1, zap104 and skn7 , and the mating control gene as the antimicrobial target gene by increasing expression is hlh1 , hap2, skn7 and gat1 It provides a method for screening an antimicrobial agent, characterized in that any one gene selected from the group consisting of.

In the above embodiment, the group consisting of 1M to 1.5M sodium chloride (NaCl), 1M to 1.5M potassium chloride (KCl), and 2M sorbitol as an antimicrobial target gene that regulates the response to external stress by reducing expression As an antimicrobial target gene against osmotic stress induced by any one selected from, the antimicrobial target genes against osmotic stress caused by sodium chloride are rim101, skn7, ada2, fzc42, hcm1, gat7, bzp2, hob1, hap2, hob6, aro8001, It is any one gene selected from the group consisting of pan1, fzc34, bap1, fzc19, fzc51, fzc43, fzc13, gat5 and met32 , and the antimicrobial target genes against osmotic stress caused by potassium chloride are bzp2, hob2, nrg1, hap2, ada2, fzc6 , yrm103, fzc44, fzc32, hob1, rim101, bzp4 and fzc35 is any one gene selected from the group consisting of, and the antimicrobial agent target gene against osmotic stress caused by sorbitol is any one gene selected from the group consisting of bzp2, hob1 and fzc6 ego; 2.5 mM to 3.5 mM hydrogen peroxide (Hydrogen Peroxdie, H ₂ O ₂ ), 0.7 mM to 0.8 mM tert-butyl hydroperoxide (TH), 0.02 mM to 0.03 mM Menadione and 2 In the antimicrobial target gene for oxidative stress stress induced by any one selected from the group consisting of mM to 3 mM diamide (DA, Diamide), the antimicrobial target genes for oxidative stress caused by hydrogen peroxide are bap1, sre1, usv101, fzc50, fzc31, hob1, ada2, cuf1, gat204, ste12, fzc9, gat1, fzc21, nrg1, bzp2, gat5, pan1, met32, fzc4, rim101, hob6, fzc13, hlh1, fzc46, sip5, fzc27, hob5, fzc4 sp1(crz1), fzc22, bzp3, liv1, miz1, and gat201 are any one gene selected from the group consisting of, and tertiary-butyl hydrogen peroxide-induced antimicrobial target genes for oxidative stress are sre1, ada2, rim101, bap2, bap1, usv101, fzc31, hob1, fzc34, ecm22, fzc15, fzc44, zfc4, fzc49, yrm103, fzc21, zfc2, gat5, pan1, met32, hob4, liv1, mwc2, skn7, hcm1, fzc51, fzc1, grf1, fzc1 bzp5, gat8, clr1, hlh2, rlm1, fzc6, asg1, hob2, and a zap103 any one gene selected from the group consisting of antibacterial agents target gene to oxidative stress due to the menadione is bap1, fzc37, usv101, nrg1, bzp2 , fzc4, fzc34, fzc35, hel2, ecm22, fzc6, gat6, jjj1, fzc44, fzc3 and fzc2 It is any one gene selected from the group consisting of 6, and the antimicrobial target genes against oxidative stress caused by diamides are bap1, hob1, bap2, hap2, pip2, bzp5, hsf2, sre1, fzc21, zfc2, fzc31, bzp2, gat5, Any one selected from the group consisting of pan1, met32, fzc4, fzc34, hob6, hlh1, fzc46, sip402, fzc27, miz1, fzc19, hlh3, fkh2, mln1, gat6, fap1, fzc8, fzc49, fzc3, fzc30, rum1 and fzc38 Is the gene of; Antimicrobial agent against endoplasmic reticulum stress caused by vesicle stress stress induced by 0.3 ㎍/mL tunicamycin (TM, tunicamycin) or 20 mM dithiothreitol (DTT) The target gene is any selected from the group consisting of bzp1 (hxl1), sre1, hlh1, bzp3, pip2, rlm1, met32, ste12, rim101, sp1 (crz1), fzc21, gat7, mln1, fzc2, fzc44, liv4, fzc40 and fzc38 . It is one gene, and the antimicrobial target genes for endoplasmic reticulum stress by dithiothreitol are bzp1 (hxl1), sre1, bzp2, cuf1, hob1, clr1, ada2, rlm1, gat5, hap2, nrg1, usv101, fzc31, gat201, It is any one gene selected from the group consisting of hlh2, apn2, fzc25, and ddt1 , and in 0.03% to 0.06% methyl methanesulfonate (MMS) or 50 mM to 100 mM hydroxyurea (HU, hydroxyurea). In the antimicrobial target gene for genotoxic stress induced by stress, the antimicrobial target genes for genotoxic stress by methyl methanesulfonate are bzp1(hxl1), fzc6, hob1, sre1, gat5, gat6, miz1, bzp2, jjj1, It is any one gene selected from the group consisting of fzc40, fzc38, fzc4, hcm101, fzc1 and apn2 , and the antimicrobial target genes for genotoxic stress caused by hydroxyurea are hob1, sre1, gat5, gat6, mbs1, skn7, ada2, bzp1 (hxl1), fzc6, bzp2, jjj1, hcm1, nrg1, and hlh2 . Cell wall or cell membrane stress induced by any one selected from the group consisting of CFW, calcofluor white), 0.8% to 1% congo red (CR) and 0.03% sodium dodecyl sulfate (SDS) In the antimicrobial target gene for stress, the antimicrobial target gene for cell wall or cell membrane stress by CFW is any selected from the group consisting of bzp1(hxl1), sp1(crz1), hob1, hap2, bzp2, nrg1, bap2, rim101 and pip2. One gene, and the gene that regulates responsiveness to cell wall or cell membrane stress by CR is any one selected from the group consisting of sp1(crz1), hob1, bzp1(hxl1), cuf1, hlh3, hap2, bzp2, nrg1, bap2, and rim101. Sp1 (crz1), hob1, sre1, pip2, fzc21, gat7, hob3, usv101, gat201, fzc7, asg1, rum1, hap2, bzp2, nrg1, cuf1, gat5, gat6, jjj1, pan1, bzp3, rlm1, bap1, clr1, zfc4, clr4, gat1, fzc31, hob5, asg101, ert1, ecm22, zfc6, bzp5, sxi1 alpha, fap1, sip4, rds2 and fzc30, fzc26 and fzc30 Any one gene selected from the group consisting of, and 20 M to 30 M of cadmium sulfate (CdSO ₄ ) Antimicrobial target genes for heavy metal stress stress are cuf1, hap2, fzc6, skn7, fzc37, bzp2, gat5, yox101, mln1, pip2, hcm1, hob6, fzc46, hob5, mbs2, fzc35, aro8001, fzc19, fzc51, aro80, ccd4, fzc47, bzp4 , fap1, fzc8, pip201, gln3, yrm101, zfc8, hob7, rum1 and fzc10 It provides a method for screening an antimicrobial agent, characterized in that any one gene selected from the group consisting of.

In the above embodiment, 1 M to 1.5 M sodium chloride (NaCl), 1 M to 1.5 M potassium chloride (KCl) and 2 M sorbitol as an antimicrobial target gene that regulates the response to external stress by increasing expression In the antimicrobial target gene against osmotic stress induced by any one selected from the group consisting of, the antimicrobial target gene against the osmotic stress caused by sodium chloride is any one gene selected from the group consisting of hlh3, hel2 and cuf1, and by potassium chloride. The antimicrobial target gene for osmotic stress is either fzc36 or yrm103 , the antimicrobial target gene for osmotic stress by sorbitol is fzc36 , 2.5 mM to 3.5 mM hydrogen peroxide (Hydrogen Peroxdie, H ₂ O ₂ ), 0.7 Any selected from the group consisting of mM to 0.8 mM tert-butyl hydroperoxide (TH), menadione of 0.02 mM to 0.03 mM, and diamide (DA, Diamide) of 2 mM to 3 mM In the antimicrobial target gene for oxidative stress caused by one, the antimicrobial target gene for oxidative stress caused by hydrogen peroxide is any one gene selected from the group consisting of fzc45, asg101, mbs2, fzc35, bwc2, fzc7 and znf2, The antimicrobial target gene for oxidative stress caused by tertiary-butyl hydrogen peroxide is any one gene selected from the group consisting of fzc33, fap1, clr3 and ddt1 , and the antimicrobial target genes for oxidative stress caused by menadione are zfc2, fzc50, cuf1. , hap2, and sip4 , and the antimicrobial target genes against oxidative stress caused by diamides are fzc50, sip4, pip201, nrg1, gat1, znf2 , asg101, skn7, gat7, jjj1, hlh5, fzc26, and any one gene selected from the group consisting of fzc20; Antimicrobial agent against endoplasmic reticulum stress induced by 0.3 μg/ml tunicamycin (TM, tunicamycin) or 20 mM dithiothreitol (DTT) antimicrobial agent against endoplasmic reticulum stress caused by the tunicamycin in the target gene Target genes are bzp2, nrg1, hap2, cuf1, mbs1, ppr1, fzc6, skn7, zfc2, hob1, gat5, clr1, bap1, bwc2, hcm1, hel2, gat6, jjj1, hob3, zfc4, zfc3 and clr4 from the group consisting of Any one selected gene, and the antimicrobial target gene for endoplasmic reticulum stress by dithiothreitol is yap4, hlh1, bzp3, pip2, pan1, mbs1, met32, gat1, fkh2, fzc11, gat203, sip401, stb4 and fzc20 . It is any one gene selected from the group, and for genotoxic stress stress caused by 0.03% to 0.06% methyl methanesulfonate (MMS) or 50 mM to 100 mM hydroxyurea (HU). In the antimicrobial target gene, the antimicrobial target gene against genotoxic stress by methyl methanesulfonate is yox1, and the antimicrobial target gene against genotoxic stress by hydroxyurea is fzc20, and 3 mg/ml to 5 mg/ml of Calco Cell wall caused by any one selected from the group consisting of fluorine white (CFW), 0.8% to 1% congo red (CR) and 0.03% sodium dodecyl sulfate (SDS) Or in the antimicrobial target gene for cell membrane stress stress, the antimicrobial target gene for cell wall or cell membrane stress by CFW is any one of fzc9 or grf1 and is caused by SDS. The antimicrobial target gene for cell wall or cell membrane stress is any one gene selected from the group consisting of fzc6, fzc1, Zfc1, hsf3, bwc2, skn7, fzc50, fzc22, fzc51 and fzc8; Antimicrobial target genes for heavy metal stress stress by 20 μM to 30 μM cadmium sulfate (CdSO ₄ ) are bzp1 (hxl1), gat201, znf2, sip4, rds2, sre1, gat7, rlm1, clr1, zfc3, ada2, gat204, It provides an antibacterial agent screening method, characterized in that it is any one gene selected from the group consisting of fzc7, asg101, atf1, hlh2, fzc39, and hsf3.

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.

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.

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.

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 is an antimicrobial agent target gene of a 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 is a target gene of an antimicrobial agent of Cryptococcus neoformans.

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.

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. .

Example

Creation of transcription factor mutant (TF mutant)

1.1 Selection of transcription factors targeting transcription factor variants

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).

List of Cryptococcus neoformans transcription factors predicted based on DNA-binding domain database No. H99 ID TF domains Gene name
(Gene name)
One
02566
"Winged helix" DNA-binding domain / Fork head domain
FKH2
2
00791
Helix-loop-helix DNA-binding domain
HLH1
3
01069
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC11
4
07464
APSES domain / Ankyrin repeat#2
MBS1
5
03401
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT203
6
04588
Fungal Zn(2)-Cys(6) binuclear cluster domain
ERT1
7
00828
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
SIP401
8
03561
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC33
9
06762
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT204
10
06276
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
CEP3
11
05785
Fungal Zn(2)-Cys(6) binuclear cluster domain
STB4
12
03132
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC5
13
01438
KilA-N domain / Ankyrin repeats (many copies)
MBS2
14
07593
bZIP transcription factor / Domain of unknown function (DUF3425)
YAP4
15
04837
Helix-loop-helix DNA-binding domain
MLN1
16
05093
Homeobox domain
HOB6
17
05642
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC37
18
05431
Zinc-finger double domain / C2H2-type zinc finger
RIM101
19
04398
Fungal specific transcription factor domain
/ Fungal Zn(2)-Cys(6) binuclear cluster domain
ARO80
20
04878
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC1
21
03183
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC24
22
03710
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
ECM22
23
03279
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
CCD4
24
04637
Helix-turn-helix/lambda repressor-like DNA-binding domains
MBF1
25
06425
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
PPR1
26
04345
Fungal Zn(2)-Cys(6) binuclear cluster domain
ARO8001
27
04184
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC47
28
02774
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
MAL13
29
00670
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC12
30
00068
Zinc-finger double domain / C2H2-type zinc finger
MET32
31
05010
C2H2-type zinc finger
ZFC7
32
04090
bZIP transcription factor / Basic region leucine zipper
ATF1
33
06134
bZIP transcription factor / Basic region leucine zipper
BZP1(HXL1)
34
04630
bZIP transcription factor / Basic region leucine zipper
BAP2
35
07901
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC29
36
01173
Beta-trefoil DNA-binding domain / p53-like transcription factors / DNA-binding protein LAG-1 (CSL)
PAN1
37
03115
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC46
38
07924
SRF-type transcription factor (DNA-binding and dimerisation
MCM1
39
07435
CCAAT-binding transcription factor (CBF-B/NF-YA) subunit B
HAP2
40
02555
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
SIP402
41
04594
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC27
42
06188
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC15
43
05170
Fungal Zn(2)-Cys(6) binuclear cluster domain
PIP2
44
02241
Homeodomain-like / Helix-turn-helix domain
HOB5
45
06921
Homeobox domain
HOB4
46
05186
GRF zinc finger
GRF1
47
00896
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC34
48
00039
C2H2-type zinc finger
ZFC6
49
07940
Basic region leucine zipper / bZIP transcription factor
BZP5
50
06814
Homeobox KN domain
SXI1alpha
51
01454
STE like transcription factor / Zinc-finger double domain
/ C2H2-type zinc finger
STE12
52
03527
C2H2-type zinc finger / C3HC4-type zinc finger
HEL2
53
05255
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC2
54
05112
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC42
55
01883
Glucocorticoid receptor-like (DNA-binding domain) / GATA zinc finger
GAT8
56
04353
C2H2-type zinc finger
CLR1
57
05375
Helix-loop-helix DNA-binding domain
HLH2
58
03998
SRF-type transcription factor (DNA-binding and dimerisation
RLM1
59
00239
bZIP transcription factor / Basic region leucine zipper
BAP1
60
06871
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC41
61
00156
C2H2-type zinc finger
SP1 (CRZ1)
62
04268
GRF zinc finger
APN2
63
05420
Zinc-finger double domain / C2H2-type zinc finger
USV101
64
00018
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC6
65
03346
bZIP transcription factor
BZP4
66
00514
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT6
67
05538
SRR1 domain / C2H2-type zinc finger / DnaJ domain
JJJ1
68
03409
"Winged helix" DNA-binding domain / CheY-like / HSF-type DNA-binding
SKN7
69
06339
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC35
70
07011
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC22
71
07506
NF-X1 type zinc finger #3 / R3H domain
FAP1
72
04807
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC8
73
02435
PYP-like sensor domain (PAS domain)
/ Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger / AT hook motif
BWC2 ( CWC2 )
74
02364
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC19
75
03116
"Winged helix" DNA-binding domain / Fork head domain
HCM1
76
02877
Zinc-finger double domain / Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC51
77
00559
bZIP transcription factor / Basic region leucine zipper
BZP3
78
03914
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC14
79
00871
bZIP transcription factor / Basic region leucine zipper
CLR3
80
06483
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC25
81
07797
Putative FMN-binding domain
CRL6
82
05019
Fungal specific transcription factor domain
FZC21
83
05380
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC44
84
04518
Fungal specific transcription factor domain / C2H2-type zinc finger
ZFC5
85
05176
Homeobox domain
HOB3
86
05861
Fork head domain
FKH101
87
00460
Helix-loop-helix DNA-binding domain
LIV1
88
02305
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC45
89
03849
Fungal specific transcription factor domain
/ Fungal Zn(2)-Cys(6) binuclear cluster domain
ASG1
90
01014
C2H2-type zinc finger
ZFC4
91
01858
Homeobox domain
HOB2
92
06719
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC49
93
04093
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
YRM103
94
04176
"Winged helix" DNA-binding domain / HSF-type DNA-binding
HSF2
95
01431
Homeobox domain
HOB1
96
07443
Helix-loop-helix DNA-binding domain
HLH4
97
04916
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC16
98
05392
Zinc-finger double domain / C2H2-type zinc finger
ZAP104
99
02322
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC17
100
04012
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC18
101
00505
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC28
102
06751
Helix-loop-helix DNA-binding domain
HLH3
103
04841
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC43
104
03212
"Winged helix" DNA-binding domain
HCM101
105
01626
Zinc finger, ZZ type / Myb-like DNA-binding domain
/ SWIRM domain
ADA2
106
03894
Fungal Zn(2)-Cys(6) binuclear cluster domain
PDR802
107
02066
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC13
108
06818
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
HAP1
109
05940
C2H2-type zinc finger
ZFC3
110
01948
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC36
111
04804
Helix-loop-helix DNA-binding domain
SRE1
112
04352
Zinc-finger double domain / C2H2-type zinc finger
ZAP103
113
03768
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC32
114
01977
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC39
115
04895
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC3
116
04583
WSTF, HB1, Itc1p, MBD9 motif 2/3 / DDT domain
DDT1
117
01973
Zinc-finger double domain / C2H2-type zinc finger
/ Fungal specific transcription factor domain
ZFC2
118
02603
Fungal specific transcription factor domain
/ C2H2-type zinc finger
ZFC1
119
04036
"Winged helix" DNA-binding domain / HSF-type DNA-binding
HSF3
120
06156
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC7
121
03431
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC48
122
07922
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC4
123
00031
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
MLR1
124
03018
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
ASG101
125
04263
Glucocorticoid receptor-like (DNA-binding domain) / Basic region leucine zipper
/ bZIP transcription factor / GATA zinc finger
BZP2
126
01708
GATA zinc finger
GAT7
127
00332
Fungal Zn(2)-Cys(6) binuclear cluster domain
SIP4
128
07724
Copper fist DNA binding domain
CUF1
129
03902
Fungal Zn(2)-Cys(6) binuclear cluster domain/PAS fold
RDS2
130
03366
Zinc-finger double domain / C2H2-type zinc finger
ZNF2
131
05222
C2H2-type zinc finger / Zinc-finger double domain
NRG1
132
05049
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
PIP201
133
02516
Helix-loop-helix DNA-binding domain
HLH5
134
04774
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC26
135
03059
Fungal Zn(2)-Cys(6) binuclear cluster domain
FZC9
136
00193
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT1
137
03336
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC50
138
03086
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC20
139
03229
Homeobox domain / Homeobox KN domain
YOX101
140
01841
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GLN3
141
02476
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
YRM101
142
05153
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger / AT hook motif
GAT5
143
02700
C2H2-type zinc finger
ZFC8
144
04586
Homeobox domain
HOB7
145
02723
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC23
146
03741
Fungal Zn(2)-Cys(6) binuclear cluster domain / AT hook motif
FZC31
147
04457
Fungal Zn(2)-Cys(6) binuclear cluster domain / AT hook motif
FZC30
148
06283
Homeodomain-like
LIV4
149
07411
C5HC2 zinc finger / PHD-finger / ARID/BRIGHT DNA binding
RUM1
150
04836
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC10
151
00841
Fungal Zn(2)-Cys(6) binuclear cluster domain
/ Fungal specific transcription factor domain
FZC40
152
06223
MIZ/SP-RING zinc finger
MIZ1
153
00830
Fungal Zn(2)-Cys(6) binuclear cluster domain / Fungal specific transcription factor domain
FZC38
154
01551
Glucocorticoid receptor-like (DNA-binding domain)
/ GATA zinc finger
GAT201
155
04908
bZIP transcription factor / Basic region leucine zipper
CLR4

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.

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 (Non-Patent Document 9, Non-Patent Document 10, and Non-Patent Document 11). Two or more independent transcription factor mutants (TF mutants) were produced for 155 transcription factors, and 322 strains were obtained (see Table 2).

Transcription factor (TF) variant division H99 lD Designated name TF class Strain informatin
One
02566
FKH2
FKH
YSB1339
2
YSB1340
3
00791
HLH1
HLH
YSB1175
4
YSB1176
5
07464
MBS1
APS
YSB488
6
YSB489
7
03401
GAT203
GAT
YSB569
8
YSB570
9
04588
ERT1
FZC
YSB693
10
YSB694
11
00828
SIP401
FZC
YSB1358
12
YSB1359
13
03561
FZC33
FZC
YSB1074
14
YSB1075
15
06762
GAT204
GAT
YSB1311
16
YSB1312
17
06276
CEP3
FZC
YSB847
18
YSB848
19
05785
STB4
FZC
YSB1013
20
YSB1014
21
01438
MBS2
APS
YSB538
22
YSB539
23
07593
YAP4
BZP
YSB1587
24
YSB1661
25
04837
MLN1
HLH
YSB1172
26
YSB1173
27
05093
HOB6
HOM
YSB1255
28
YSB1256
29
05642
FZC37
FZC
YSB1329
30
YSB1330
31
05431
RIM101
C2Z
YSB1366
32
YSB1367
33
04398
ARO80
FZC
YSB714
34
YSB715
35
04878
FZC1
FZC
YSB510
36
YSB511
37
03183
FZC24
FZC
YSB774
38
YSB775
39
03710
ECM22
FZC
YSB476
40
YSB478
41
03279
CCD4
HOM
YSB706
42
YSB707
43
04637
MBF1
HTH
YSB768
44
YSB769
45
06425
PPR1
FZC
YSB1046
46
YSB1047
47
04345
ARO8001
FZC
YSB661
48
YSB662
49
04184
FZC47
FZC
YSB1406
50
YSB1407
51
02774
MAL13
FZC
YSB506
52
YSB507
53
00670
FZC12
FZC
YSB467
54
YSB468
55
05010
ZFC7
C2Z
YSB481
56
YSB482
57
04090
ATF1
BZP
YSB676
58
YSB678
59
06134
BZP1(HXL1)
BZP
YSB723
60
YSB724
61
04630
YAP2
BZP
YSB1416
62
YSB1417
63
07901
FZC29
FZC
YSB718
64
YSB719
65
03115
FZC46
FZC
YSB1209
66
YSB1210
67
07924
MCM1
SRF
YSB1302
68
YSB1303
69
07435
HAP2
CCA
YSB1104
70
YSB1105
71
02555
SIP402
FZC
YSB529
72
YSB530
73
04594
FZC27
FZC
YSB582
74
YSB583
75
06188
FZC15
FZC
YSB646
76
YSB647
77
05170
PIP2
FZC
YSB1249
78
YSB1250
79
02241
HOB5
HOM
YSB1585
80
YSB1586
81
06921
HOB4
HOM
YSB1435
82
YSB1437
83
05186
GRF1
GRF
YSB796
84
YSB797
85
00039
ZFC6
C2Z
YSB1953
86
YSB1954
87
01454
STE12
C2Z
YSB1542
88
YSB1543
89
03527
HEL2
C2Z
YSB1382
90
YSB1383
91
05255
FZC2
FZC
YSB1050
92
YSB1051
93
01883
GAT8
GAT
YSB471
94
YSB472
95
04353
CLR1
C2Z
YSB1396
96
YSB1397
97
03998
RLM1
SRF
YSB1300
98
YSB1301
99
00239
YAP1
BZP
YSB815
100
YSB1290
101
06871
FZC41
FZC
YSB1334
102
YSB1335
103
00156
SP1 (CRZ1)
C2Z
YSB1263
104
YSB1264
105
04268
APN2
GRF
YSB1429
106
YSB1430
107
05420
USV101
C2Z
YSB1464
108
YSB1465
109
00018
FZC6
FZC
YSB1980
110
YSB1981
111
03346
BZP4
BZP
YSB1894
112
YSB1895
113
05538
JJJ1
C2Z
YSB1592
114
YSB1594
115
03409
SKN7
HSF
YSB349
116
YSB350
117
06339
FZC35
FZC
YSB1341
118
YSB1342
119
07506
FAP1
NFX
YSB813
120
YSB817
121
04807
FZC8
FZC
YSB2112
122
YSB2113
123
02435
BWC2
GAT
YSB1839
124
YSB1840
125
02364
FZC19
FZC
YSB2115
126
YSB2116
127
03116
HCM1
FKH
YSB1850
128
YSB1851
129
02877
FZC51
FZC
YSB1842
130
YSB1843
131
00559
BZP3
BZP
YSB1099
132
YSB1100
133
03914
FZC14
FZC
YSB1846
134
YSB1847
135
00871
CLR3
BZP
YSB1834
136
YSB1836
137
06483
FZC25
FZC
YSB518
138
YSB1822
139
07797
CRL6
FKH
YSB1106
140
YSB1107
141
05019
FZC21
FZC
YSB1252
142
YSB1253
143
05380
FZC44
FZC
YSB2182
144
YSB2183
145
04518
ZFC5
C2Z
YSB2177
146
YSB2178
147
05176
HOB3
HOM
YSB2001
148
YSB2002
149
05861
FKH101
FKH
YSB1855
150
YSB1856
151
00460
LIV1
HLH
YSB2211
152
YSB2212
153
02305
FZC45
FZC
YSB2221
154
YSB2222
155
03849
ASG1
FZC
YSB3013
156
YSB3014
157
01014
ZFC4
C2Z
YSB2231
158
YSB2232
159
01858
HOB2
HOM
YSB2282
160
YSB2283
161
06719
FZC49
FZC
YSB2171
162
YSB2173
163
04093
YRM103
FZC
YSB2298
164
YSB2299
165
04176
HSF2
HSF
YSB2295
166
YSB2296
167
01431
HOB1
HOM
YSB2308
168
YSB2309
169
07443
HLH4
HOM
YSB2244
170
YSB2245
171
04916
FZC16
FZC
YSB2326
172
YSB2327
173
05392
ZAP104
C2Z
YSB2134
174
YSB2135
175
02322
FZC17
FZC
YSB2250
176
YSB2251
177
04012
FZC18
FZC
YSB2320
178
YSB2321
179
00505
FZC28
FZC
YSB2337
180
YSB2338
181
06751
HLH3
HLH
YSB2329
182
YSB2330
183
04841
FZC43
FZC
YSB517
184
YSB2334
185
03212
HCM101
FKH
YSB2390
186
YSB2391
187
01626
ADA2
MYB
YSB2381
188
YSB2382
189
03894
PDR802
FZC
YSB2387
190
YSB2388
191
02066
FZC13
FZC
YSB2517
192
YSB2518
193
06818
HAP1
FZC
YSB2481
194
YSB2482
195
05940
ZFC3
C2Z
YSB2108
196
YSB2386
197
01948
FZC36
FZC
YSB2335
198
YSB2523
199
04804
SRE1
HLH
YSB2493
200
YSB2494
201
04352
ZAP103
C2Z
YSB2540
202
YSB2541
203
03768
FZC32
FZC
YSB2385
204
YSB2526
205
04895
FZC3
FZC
YSB2611
206
YSB2664
207
04583
DDT1
DDT
YSB1583
208
YSB2633
209
01973
ZFC2
C2Z
YSB2622
210
YSB2623
211
06156
FZC7
FZC
YSB2704
212
YSB2705
213
03431
FZC48
FZC
YSB2646
214
YSB2647
215
07922
FZC4
FZC
YSB2724
216
YSB2725
217
00031
MLR1
FZC
YSB2727
218
YSB2728
219
03018
ASG101
FZC
YSB2697
220
YSB2698
221
04263
BZP2
BZP
YSB2702
222
YSB2703
223
01708
GAT7
GAT
YSB2699
224
YSB2700
225
00332
SIP4
FZC
YSB2680
226
YSB2681
227
07724
CUF1
CDB
YSB2665
228
YSB2666
229
03902
RDS2
FZC
YSB1898
230
YSB1899
231
03366
ZNF2
C2Z
YSB2740
232
YSB2741
233
05222
NRG1
C2Z
YSB3096
234
YSB3097
235
05049
PIP201
FZC
YSB3099
236
YSB3100
237
02516
HLH5
HLH
YSB2609
238
YSB3059
239
04774
FZC26
FZC
YSB3084
240
YSB3085
241
03336
FZC50
FZC
YSB3131
242
YSB3132
243
03086
FZC20
FZC
YSB3128
244
YSB3129
245
03229
YOX101
HOM
YSB3134
246
YSB3136
247
01841
GLN3
GAT
YSB3154
248
YSB3155
249
02476
YRM101
FZC
YSB2997
250
YSB2998
251
05153
GAT5
GAT
YSB3033
252
YSB3034
253
02700
ZFC8
C2Z
YSB3031
254
YSB3032
255
04586
HOB7
HOM
YSB3026
256
YSB3027
257
02723
FZC23
FZC
YSB3105
258
YSB3106
259
03741
FZC31
FZC
YSB3093
260
YSB3094
261
04457
FZC30
FZC
YSB2447
262
YSB2448
263
04836
FZC10
FZC
YSB3083
264
YSB3368
265
06223
MIZ1
MIZ
YSB2133
266
YSB3366
267
01551
GAT201
GAT
YSB3300
268
YSB3301
269
04908
CLR4
BZP
YSB3282
270
YSB3283
271
07940
BZP5
BZP
YSB1474
272
YSB1475
273
05112
FZC42
FZC
YSB687
274
YSB690
275
00193
GAT1
GAT
YSB2972
276
YSB2973
277
06814
SXI1alpha
HOM
YSB1390
278
YSB1391
279
YSB1392
280
00896
FZC34
FZC
YSB501
281
YSB2979
282
07411
RUM1
PHZ
YSB3164
283
YSB3747
284
00830
FZC38
FZC
YSB777
285
YSB3791
286
01069
FZC11
FZC
YSB845
287
YSB846
288
YSB2983
289
03132
FZC5
FZC
YSB1400
290
YSB1401
291
YSB1404
292
00068
MET32
C2Z
YSB1178
293
YSB1179
294
YSB1180
295
01173
PAN1
P53
YSB1181
296
YSB1182
297
YSB1183
298
00514
GAT6
GAT
YSB1384
299
YSB1385
300
YSB1386
301
07011
FZC22
FZC
YSB1688
302
YSB1689
303
YSB2974
304
02603
ZFC1
C2Z
YSB2573
305
YSB2574
306
YSB2575
307
04036
HSF3
HSF
YSB2527
308
YSB2528
309
YSB2529
310
03059
FZC9
FZC
YSB2984
311
YSB3266
312
YSB3267
313
06283
LIV4
MYB
YSB2089
314
YSB3755
315
YSB3756
316
00841
FZC40
FZC
YSB3088
317
YSB3758
318
01977
FZC39
FZC
YSB1820
319
YSB2621
320
05375
HLH2
HLH
YSB1147
321
YSB1148
322
YSB1149

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. Known transcription factors including RIM101 , ADA2 , CUF1 , SXL1 , SP-1/CRZ1 , NRG1 , STE12 , BWC2 , SRE1 , 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).

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 Creation of transcription factor variants

A transcription factor deletion variant (hereinafter, TFKO) of the Cryptococcus neoformans strain was prepared against the background of the C. neoformans serotype A H99S strain. The gene disruption cassette containing 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 (Non-Patent Document 15 and Non-Patent Document 16). During the overlapping PCR process, the 5'and 3'adjacent regions of the transcription factor gene were amplified using primers L1 and R1, primers L2 and R2 (see Table 1) for H99 genomic DNA, respectively, in the first round of PCR. . Primer M13Fe (M13 forward extension) and primer M13Re (M13 reverse extension) were used to amplify ^{a dominant selection marker (NAT r) comprising a unique resistance marker sequence.} In the second round of PCR, the transcription factor gene-disrupting cassette was constructed by performing overlapping PCR using primers L1 and R2 using a 1-round PCR product as a template. In the double joint PCR method, 5'and 3'adjacent regions of the transcription factor gene were amplified using H99 genomic DNA and primer pairs L1/L2 and R1/R2, respectively, in the first round of PCR. The 5'and 3'regions of the NAT split marker were amplified using primers M13Fe, NSL, primers M13Re, and NSR against pNATSTM (obtained from Duke University's Joeseph Heitman laboratory) containing a unique signature-tag sequence. The amplified gene-disrupting cassette was combined with 600 ug of microtransport gold beads (0.6 um, Bio-Rad), and treated with 10 µl of 2.5M calcium chloride and 2 µl of 1 M spermidine. The gold beads to which the gene-disrupting cassette was bound were introduced into the H99S strain (obtained from Joeseph Heitman Laboratory, Duke University) using a biolistic transformation device (Non-Patent Document 17). After incubation for 4 hours required for recovery, the cells were scraped and transferred onto a yeast extract-peptone dextrose (hereinafter, YPD) medium containing norcerosine (100 µg/ml), and thereafter 3 to 5 Incubated for days. Stable norsetrosine-resistant transformants were screened by performing diagnostic PCR using the primer sets listed in Table 3 and Sequence Listing.

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.

Construction of primer sets for genes Gene name Primer name Primer details FZC5






L1 CNAG_03132 5'flanking region primer 1 L2 CNAG_03132 5'flanking region primer 2 R1 CNAG_03132 3'flanking region primer 1 R2 CNAG_03132 3'flanking region primer 2 SO1 CNAG_03132 diagnostic screening primer, pairing with B79 PO2 CNAG_03132 Southern blot probe primer STM NAT#5 STM primer STM common STM common primer

Phenotypic profiling

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.

All data on the phenome are publicly available and available in the Cryptococcus neoformans transcription factor database (http://tf.cryptococcus.org).

2.1 Genotyping

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 pH 8 and allowed to stand for 45 minutes. The cut genomic DNA was transferred to a nylon membrane using 10-fold Saline-Sodium Citrate (SSC) buffer, and was fixed by exposure to ultraviolet light of ^{1200 J/m 2.} The membrane was gene-specific and radioactive using a modified church hybridization buffer (1 mM EDTA, 0.25 M Na ₂ HPO ₄ , 1% hydrolyzed casein, 7% SDS, 6% H ₃ PO _{4 ).} Hybridization with the labeled probe. The membrane was washed with Wash Buffer 1 (with 2X SSC and 0.1% SDS) and Wash Buffer 2 (with 1X SSC and 0.1% SDS) for 15 minutes. Thereafter, the membrane was exposed to an automation film for 1 day.

Transcription factor deletion variants and list of restriction enzymes used division

H99 lD
Designated name
Restriction enzyme cut
One
02566
FKH2
BamHI
2
00791
HLH1
BglII
3
01069
FZC11
SphI
4
07464
MBS1
SphI
5
03401
GAT203
HindIII
6
04588
ERT1
EcoRV
7
00828
SIP401
EcoRI
8
03561
FZC33
EcoRV
9
06762
GAT204
KpnI
10
06276
CEP3
SphI
11
05785
STB4
BamHI
12
03132
FZC5
PstI
13
01438
MBS2
EcoRI
14
07593
YAP4
SmaI
15
04837
MLN1
SmaI
16
05093
HOB6
KpnI
17
05642
FZC37
XmaI
18
05431
RIM101
ClaI
19
04398
ARO80
HincII
20
04878
FZC1
EcoRI
21
03183
FZC24
EcoRV
22
03710
ECM22
HindIII
23
03279
CCD4
HindIII
24
04637
MBF1
HindIII
25
06425
PPR1
EcoRV
26
04345
ARO8001
BamHI
27
04184
FZC47
BglII
28
02774
MAL13
EcoRV
29
00670
FZC12
StyI
30
00068
MET32
AfeI
31
05010
ZFC7
ScaI
32
04090
ATF1
PstI
33
06134
BZP1(HXL1)
BamHI
34
04630
YAP2
HindIII
35
07901
FZC29
EcoRV
36
01173
PAN1
SalI
37
03115
FZC46
SacI
38
07924
MCM1
XbaI
39
07435
HAP2
SalI
40
02555
SIP402
MfeI
41
04594
FZC27
HindIII
42
06188
FZC15
BamHI
43
05170
PIP2
BamHI
44
02241
HOB5
PstI
45
06921
HOB4
KpnI
46
05186
GRF1
EcoRI
47
00896
FZC34
HindIII
48
00039
ZFC6
KpnI
49
07940
BZP5
BamHI
50
06814
SXI1alpha
SacI
51
01454
STE12
BamHI
52
03527
HEL2
BamHI
53
05255
FZC2
HindIII
54
05112
FZC42
BglII, EcoRV
55
01883
GAT8
SacI
56
04353
CLR1
SalI
57
05375
HLH2
HindIII
58
03998
RLM1
StyI
59
00239
YAP1
BamHI
60
06871
FZC41
BamHI, XbaI
61
00156
SP1 (CRZ1)
EcoRV
62
04268
APN2
EcoRV
63
05420
USV101
SphI
64
00018
FZC6
SphI
65
03346
BZP4
PstI
66
00514
GAT6
EcoRV
67
05538
JJJ1
SphI
68
03409
SKN7
BamHI
69
06339
FZC35
HindIII
70
07011
FZC22
PstI
71
07506
FAP1
HindIII
72
04807
FZC8
PstI
73
02435
BWC2
HindIII
74
02364
FZC19
BamHI
75
03116
HCM1
SphI
76
02877
FZC51
SphI
77
00559
BZP3
SphI
78
03914
FZC14
PstI
79
00871
CLR3
XbaI
80
06483
FZC25
PstI
81
07797
CRL6
EcoRI
82
05019
FZC21
PstI
83
05380
FZC44
SacI
84
04518
ZFC5
EcoRI
85
05176
HOB3
XhoI
86
05861
FKH101
EcoRV
87
00460
LIV1
SphI
88
02305
FZC45
EcoRV
89
03849
ASG1
KpnI
90
01014
ZFC4
SalI
91
01858
HOB2
SphI
92
06719
FZC49
PstI
93
04093
YRM103
SphI
94
04176
HSF2
KpnI
95
01431
HOB1
HindIII
96
07443
HLH4
SphI
97
04916
FZC16
EcoRI
98
05392
ZAP104
SalI
99
02322
FZC17
XbaI
100
04012
FZC18
HindIII
101
00505
FZC28
SmaI
102
06751
HLH3
SalI
103
04841
FZC43
XbaI
104
03212
HCM101
SacI, SalI
105
01626
ADA2
BamHI
106
03894
PDR802
SphI
107
02066
FZC13
BamHI
108
06818
HAP1
HindIII
109
05940
ZFC3
EcoRI
110
01948
FZC36
XhoI
111
04804
SRE1
BamHI
112
04352
ZAP103
SacI, SalI
113
03768
FZC32
BamHI
114
01977
FZC39
KpnI
115
04895
FZC3
KpnI
116
04583
DDT1
SphI
117
01973
ZFC2
SphI
118
02603
ZFC1
HindIII
119
04036
HSF3
SphI
120
06156
FZC7
EcoRI
121
03431
FZC48
EcoRV
122
07922
FZC4
EcoRV
123
00031
MLR1
PstI, XmaI
124
03018
ASG101
XhoI
125
04263
BZP2
EcoRV
126
01708
GAT7
SacI
127
00332
SIP4
PstI
128
07724
CUF1
KpnI
129
03902
RDS2
EcoRV, EcoRI
130
03366
ZNF2
EcoRV
131
05222
NRG1
EcoRV
132
05049
PIP201
BamHI
133
02516
HLH5
BamHI
134
04774
FZC26
PstI
135
03059
FZC9
EcoRV
136
00193
GAT1
SphI
137
03336
FZC50
HindIII
138
03086
FZC20
KpnI
139
03229
YOX101
XbaI
140
01841
GLN3
EcoRI
141
02476
YRM101
XhoI
142
05153
GAT5
SalI
143
02700
ZFC8
EcoRV
144
04586
HOB7
SphI
145
02723
FZC23
EcoRV
146
03741
FZC31
ClaI
147
04457
FZC30
BamHI
148
06283
LIV4
EcoRV
149
07411
RUM1
EcoRV
150
04836
FZC10
EcoRV
151
00841
FZC40
EcoRI
152
06223
MIZ1
EcoRV
153
00830
FZC38
SphI
154
01551
GAT201
XbaI
155
04908
CLR4
HindIII

2.2 Gene expression analysis

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.

Transcription factor (TF) that regulates growth

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). Because cuf1 wild-type growth is restored by external CuSO ₄ administration, the growth deficiency of the cuf1 mutant is due to impaired copper absorption.

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 , CRZ1 , 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. Conversely, 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.

A transcription factor that regulates mating

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).

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 (pH 5, 1L standard: V8 juice 50 ml (Campbell), KH2PO4 (Bioshop, PPM302) 0.5g, Agar (Bioshop, AGR001.500) 40g) were spotted, and cultured by blocking light at room temperature for 1 to 2 weeks. Filamentous growth was observed weekly and photographed using an Olympus BX51 microscope equipped with a SPOT Insight Digital Camera (Diagnostic Instrument Inc.).

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.

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, SKN 7, which promotes mating, is essential for pheromone gene expression and cell fusion (Figs. 6b and 6c), suggesting that SKN 7 is involved in the late stages of mating (Fig. 6d). In the analysis according to the present invention, it was suggested that 34 transcription factors are involved in mating (FIG. 7a-positive regulator, FIG. 7b-negative regulator).

Transcription factors that regulate the production of toxic factors

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 Capsule production

Capsule production was measured by qualitative and quantitative methods as described in prior literature (Non-Patent Literature 20 and Non-Patent Literature 21). Cells were grown for 16 hours at 30°C in liquid YPD medium, spotted with DME (Dulbeccos modified Eagle) solid medium (Gibco, #12100-046), and then cultured at 37°C for 2 days. After incubation, the cells were scraped from DME solid medium and washed with PBS (Phosphate-Buffed Saline). Capsule production was qualitatively measured by staining with India ink (Bactidrop; Remel) and visualizing using an Olympus BX51 microscope equipped with a spot observation digital camera (Diagnostic Instrument Inc.). For qualitative analysis, cells collected from DME solid medium were fixed with 10% formalin, and the same number of cells (2.5×10 ⁷ cells/ml) were loaded into hematocrit capillaries. The hematocrit capillary was placed vertically so that the cells were compressed by gravity for 10 days. The compressed cell volume ratio was measured by calculating the length ratio of the compressed cell volume phase to the length of the total volume phase (cell + medium). The relative compressed cell volume of each variant was measured by calculating the ratio of the volume ratio of the compressed cells of the variant to the volume ratio of the compressed cells of the wild type.

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).

In the present invention, it was confirmed that 49 transcription factors were involved in capsule production (FIG. 8a, 8b-negative regulator 29, 8c-positive regulator 20). Conventionally known capsule-regulating transcription factors Atf1 (non-patent document 10), Mbs1 (non-patent document 11), Gat201 (non-patent document 12) and Ada2 (non-patent document 22) are included. In addition to these capsule-regulated transcription factors, the present inventors have identified several novel capsule-regulated transcription factors. Compared to the results observed in the previously known gat201Δ and ada2Δ mutants, the zap104Δ , bap1Δ and rds2Δ mutants significantly reduced capsule production (FIG. 8A). Thus, Ada2, Gat201, Zap104, Bap1 and Rds2 were predicted as major positive regulators in capsule production, along with other 15 positive regulators. Conversely, deletion of HOB7, CLR3 and FZC51 significantly improved capsule production (FIG. 8A ), which, along with other 26 negative modulators, was predicted to be a major negative regulator of capsule production.

5.2 Measurement of melanin production and analysis of LAC1 expression

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-positive regulator 11 kinds, 9b- negative regulator 16 kinds). A small number of transcription factors including Cuf1, Ste12, Mbs1, Skn7, and Atf1 are previously published transcription factors (Non-Patent Document 10, Non-Patent Document 11, Non-Patent Document 23, Non-Patent Document 24, and Non-Patent Document 25 ). In addition to these results, it was confirmed that the fzc8Δ , hob1Δ and bzp4Δ mutants significantly reduced melanin production, which was similar to the results of the cuf1Δ mutant (Fig. 9a).

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 Urease production

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). .

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.

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.

Transcription factors affecting Cryptococcus neoformans toxicity

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 Insect test

Insect-based toxicity test was conducted with some modifications of the existing method (Non-Patent Document 21). Fifteen (Galleria mellonella ) larvae (weight: 250±50 mg) of the final young larval stage that arrived within 7 days from the date of shipment (vanderhorst Inc., St Marys, OH, USA) were randomly sorted into each group. After that, a beehive 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.

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 Document 9, Non-Patent Document 12, Non-Patent Document 22, Non-Patent Document 27, Non-Patent Document 28, Non-Patent Document 29, Non-Patent Document 30, Non-Patent Document 31, and Non-Patent Document 32). These results suggested that the insect model and the murine animal model have a profound relationship in Cryptococcus neoformans pathogenicity. In addition to mutations in known transcription factors, mutations in HOB1 , BZP2 , USV101 , BAP1 , ZFC2 , FZC1 , FZC50 , and FZC31 significantly reduced the toxicity of Cryptococcus neoformans (FIG. 11). In the present invention, 17 transcription factor genes related to the toxicity of Cryptococcus neoformans were verified using an insect host model (FIG. 11).

6.2 Animal testing

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.

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 STM#282 and STM#169 sequences were used as toxic control and non-toxic control strains, respectively (Non-Patent Document 9 and Non-Patent Document 33). Therefore, the number of transcription factors (TF) exhibiting the same signature mark is less than 4. In the present invention, four toxicity tests based on STM were conducted. In order to obtain the input transcription factor genomic DNA library, the transcription factor variants registered in the pool were serially diluted 10-fold, plated on YPD medium, incubated at 30° C. for 3 days, scraped and collected, and used for genomic DNA extraction. . The output transcription factor genomic DNA library was performed as follows. 7-week-old female A/Jcr mice (Jacson Laboratory) anesthetized by intraperitoneal injection of avertin (2,2,2-tribromoethanol) were registered transcription factor variants 5×10 ⁵ cells (50 μl). Volume) was infected by intranasal inhalation and sacrificed by overdosing avertin 15 days after infection. For each test, 5 mice were used in the present invention. Two independent variants for each transcription factor were tested in a separate STM set test. Mouse lungs were cut and homogenized using PBS. Each lung-tissue lysate was spread on YPD medium containing 100 μg/ml chloroamphenicol and incubated at 30° C. for 3 days. After incubation, it was scraped and collected to extract the output genomic DNA. Input and output genomic DNA were extracted according to the CTAB method (Non-Patent Document 21). Quantitative PCR analysis was performed using the various label-specific primers described in Table S1 and MyiQ2 real-time PCR detection system (Bio-Rad). The STM value (Log2 [output/input]) was calculated according to the method described in the prior literature (Non-Patent Literature 12, Non-Patent Literature 34).

6.3 Animal test results

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 Document 9, Non-Patent Document 12, Non-Patent Document 29, and Non-Patent Document 30). The gat201Δ variant and the pdr802Δ variant exhibited significantly reduced STM values (-11.125 and -7.212, respectively) compared to the existing literature (Non-Patent Document 12). Similarly, STM figures zap104Δ liv1Δ variants and mutants was also reduced (-5.528 and -3.875, respectively). On the other hand, the zap103Δ variant had very high toxicity as described in the existing literature (Non-Patent Document 12) (STM value = 2.51). Additionally, hxl1Δ mutant, nrg1Δ mutant, and bwc2Δ mutant also showed significantly reduced STM values. Of the 40 transcription factors identified by STM analysis, 11 transcription factors ( GAT201 , PDR802 , HXL1 , BWC2 , NRG1 , FZC1 , HOB1 , USV101 , ZFC2 , SRE1 and FZC31 ) were reconfirmed using an insect model. The toxicity test data using the insect model was statistically highly correlated with the STM-based infectivity data from the murine animal model based on the Pearson Correlation Coefficient (PCC) analysis (FIG. 12). Of the 26 novel virulence-related variants of transcription factors screened using only the STM-based murine model, the fzc31Δ variant and the ddt1Δ variant showed significantly reduced STM levels (-4.328 and -4.832, respectively). In the phenome database according to the present invention, the fzc31Δ mutant showed increased sensitivity to osmotic pressure, oxidation, and cell membrane stress, and it was predicted that these sensitivities were aggregated to affect toxicity. Conversely, one notable phenotype observed in the ddt1Δ variant is weak dithiothreitol (DTT) susceptibility, and since other DTT-reactive transcription factor variants are as toxic as the wild-type strain, the ddt1Δ mutation was not associated with a decrease in the survival of the variant in the lung. Was predicted.

Antimicrobial resistance and susceptibility test

* 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.

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.

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).

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 (Non-Patent Document 27 and Non-Patent Document 28). Since Mbs1 negatively regulates basic ERG11 expression, deletion of Mbs1 in Cryptococcus neoformans increases azole resistance but decreases polyene resistance (Non-Patent Document 11). Other transcription factors related to ERG11 expression were examined by measuring ERG11 expression in the transcription factor variants under sterol-sufficient conditions and sterol-deficient conditions (FIG. 14B). To analyze the expression of ERG2 , ERG3 , ERG5 , ERG11 , and ERG25 , wild-type, sre1 , and hob1 were incubated overnight in liquid YPD medium. The culture solution was again inoculated into 100 ml of fresh 30° C. YPD medium, and cultured _{until the OD 600} value of the culture became approximately 1.0. To prepare a zero-time sample, 50 ml of the cell culture solution was sampled, and the remaining culture was treated with fluconazole (final concentration: 10 μg/ml) for 90 minutes. Total RNA was extracted using trizol, and cDNA was synthesized using M-MuLV reverse transcription polymerase (Thermo scientific). Northern blot analysis was performed using each ERG gene-specific probe amplified using ERG gene-specific primers for total RNA in fluconazole-treated or untreated cells: primers; B5789 (5'-CAAGAAATGGAGCGTGAG-3') and B5790 (5'-CAGTGTTGTAAAGCGTGATG-3') for ERG2; B1720 (5'-ATCCCTTTTCACCGTCGCTC-3') and B1721 (5'-CGTCGTGGATGAGAATAGTCC-3') for ERG3; B671 (5'-GTTTTGTTGCCTGAGAACTGGG-3') and B674 (5'-ATCACTCAACTCGGTCCTCTCGTG-3') for ERG5; B678 (5'-TTCAGGGAACTTGGGAACAGC-3') and B1598 (5'-CAGGAGCAGAAACAAAGC-3') for ERG11; B1718 (5'-TGACCGCCTGTAGATTGTC-3') and B1719 (5'-TAGTCCCACCACCTGAAAC-3') for ERG25. Quantitative real-time PCR was performed using MyiQ2 real-time PCR detection system (Bio Rad) and respective gene-specific primers. Primers: B5789 (5'-CAAGAAATGGAGCGTGAG-3') and B6838 (5'-GGGAGGGTCGAGGATGTAGA-3') for ERG2; B1720 (5'-ATCCCTTTTCACCGTCGCTC-3') and B6838 (5'-GGGAGGGTCGAGGATGTAGA-3') for ERG3; B671 (5'-GTTTTGTTGCCTGAGAACTGGG-3') and B672 (5'-GTAGATACTGAGAGCCTGCTTGGTG-3') for ERG5; B677 (5'-AATCTCCTTACCAGCCATTCGG-3') and B678 (5'-TTCAGGGAACTTGGGAACAGC-3') for ERG11; B2695 (5'-TCGTCTTTGGCAAGCAGTC-3') and B6840 (5'-GAAGTCGTGGTGGTCAGCA-3') for ERG25; B679 (5'-CGCCCTTGCTCCTTCTTCTATG-3') and B680 (5'-TACTCGTCGTATTCGCTCTTCG-3') for ACT1. As predicted, the basal and induction levels of ERG11 were much lower in the sre1 variant than in the wild-type strain. Surprisingly, the HOB1 deletion significantly increased the basal expression level of ERG11. Expression of ERG2 , ERG3 , ERG5 and ERG25 in wild-type strains, hob1Δ strains and sre1Δ strains under sterol-sufficient conditions and sterol-deficient conditions were observed to examine whether Hob1 is involved in other ERG gene regulation. In response to sterol deficiency through fluconazole treatment, the expression of all other ERG genes was induced in the wild-type strain, but not in the sre1Δ strain (FIG. 14C). Deletion of HOB1 significantly increased the basal expression of ERG2 (Fig. 14c). Conversely, upon sterol deficiency, the induction of ERG2 , ERG3 , ERG5 , ERG11 and ERG25 in the hob1Δ mutant was reduced (Fig. 14c). Robust regulation of ERG expression is mostly lacking in the hob1Δ variant, suggesting that Hob1 is a major regulator of ergosterol gene expression (Fig. 14D, Fig. 14E). Transcription factors related to sterol biosynthesis are predicted to be related to environmental stress response and adaptation.Since pathogens face rapid environmental changes during infection, response and adaptation to environmental stress are very important in the survival and proliferation of Cryptococcus neoformans. (Non-Patent Document 3).

External stress responsiveness test

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.

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).

Functional correlation analysis

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).

Database construction for Cryptococcus neoformans transcription factors

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) (Non-Patent Document 38, Non Patent Document 39, Non-Patent Document 40, Non-Patent Document 41, Non-Patent Document 42, Non-Patent Document 43, Non-Patent Document 44, and Non-Patent Document 45).

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.

Pearson correlation coefficient network construction

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.

Korea Microorganism Conservation Center (International) KCCM51291 20150323

<110> AmtixBio Co., Ltd. <120> Novel genes for regulating the virulence of Cryptococcus neoformans and their use <130> AMTIX16P02102 <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 (34)

An antifungal agent screening method comprising the steps of:
(a) contacting a sample to be analyzed with a cell containing hob1 as an antifungal target gene;
(b) measuring the expression level of the gene; And
(c) determining that the sample is an antifungal agent when it is determined that the expression level of the gene is regulated. The method according to claim 1, wherein the target gene bzp2 regulates resistance to polyene antifungal agents, regulates fungal growth in a temperature-independent manner, and exhibits osmotic stress, oxidative stress, endoplasmic reticulum, genotoxic stress, Is a target gene for the antifungal agent. 3. The antifungal agent screening method according to claim 1 or 2, wherein the tolerance is controlled to be reduced. 3. The antifungal agent screening method according to claim 2, wherein the osmotic stress is stress caused by sodium chloride, potassium chloride, or sorbitol. 3. The antifungal agent screening method according to claim 2, wherein the oxidative stress is hydrogen peroxide, tertiary-butyl hydrogen peroxide, or a stress caused by diamide. 3. The antifungal agent screening method according to claim 2, wherein said endoplasmic reticulum stress is endoplasmic reticulum induced by dithiothreitol. 3. The antifungal compound screening method according to claim 2, wherein the genotoxic stress is stress caused by methyl methane sulfonate or hydroxyurea. 3. The antifungal compound screening method according to claim 2, wherein the cell wall or cell membrane stress is stress caused by calcofluoro white (CFW), congord (CR), or sodium dodecyl sulfate (SDS). (a) a first measuring step of bringing an antifungal agent into contact with a cell containing the antifungal target gene hob1 and measuring the expression level of the gene;
(b) a second measuring step of contacting the sample containing the antifungal drug target gene hob1 with a sample to be analyzed and the antifungal 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 adjusted to be lower than the measurement value of the first measurement step, the sample is judged to be an antimicrobial agent for coadministration &Lt; / RTI &gt; 10. The method of claim 9, wherein the antifungal target gene hob1 regulates resistance to polyene antifungal agents, regulates fungal growth in a temperature-independent manner, and is characterized by osmotic stress, oxidative stress, endoplasmic reticulum, genotoxic stress, Wherein the target gene is a target gene for stress. An antifungal agent screening method comprising the steps of:
(a) contacting a sample to be analyzed with a cell containing an antifungal target transcription factor Hob1;
(b) measuring the amount or activity of the protein; And
(c) determining that the sample is an antifungal agent when the amount or activity of the protein is measured to be decreased. 12. The method according to claim 11, wherein the cell is a fungal cell. 13. The antifungal agent screening method according to claim 12, wherein the fungus is bacillus or acacia. The antifungal agent screening method according to claim 11 or 12, wherein the antifungal agent is for preventing, treating, or preventing and treating fungal infections caused by basidiomycetes, acanthosis fungi, or acaricidal fungi and basidiomycetes. The antifungal agent screening method according to claim 11 or 12, wherein the antifungal agent is for preventing, treating, or preventing and treating crop infection or animal infection caused by bacillary, eukaryotic, or acaricidal bacteria. 15. The method according to claim 14, wherein the bacterium or acarina is selected from the group consisting of Candida sp. , Aspergillus sp. , Cryptococcus sp. , Malassezia sp. Wherein the fungus is one or more fungi selected from the group consisting of Fusarium sp. And Penicillium sp. Fungi. 16. The method of claim 15, wherein the bacterium or acarid is selected from the group consisting of Candida sp. , Aspergillus sp. , Cryptococcus sp. , Malassezia sp. Wherein the fungus is one or more fungi selected from the group consisting of Fusarium sp. And Penicillium sp. Fungi. A pharmaceutical composition for an antifungal comprising an inhibitor for an antifungal target transcription factor Hob1, wherein said inhibitor is an antibody to said protein. Wherein the inhibitor is an antisense oligonucleotide, an siRNA, an shRNA, an miRNA or a vector comprising the antisense oligonucleotide for the gene, wherein the inhibitor comprises an inhibitor for a gene encoding an antifungal target transcription factor Hob1 . The pharmaceutical composition for an antifungal agent according to claim 18 or 19, which is for preventing, treating, or preventing and treating fungal infections caused by basidiomycetes, acanthosis fungi, or acaricidal fungi and basidiomycetes. 20. The pharmaceutical composition according to claim 18 or 19 for use in combination with an azole-based or non-azole-based antifungal agent. The pharmaceutical composition according to claim 21, wherein the azole antifungal agent is at least one selected from the group consisting of fluconazole, itraconazole, voriconazole and ketoconazole. 22. The method of claim 21, wherein the non-azole based antifungal agent is selected from the group consisting of amphotericin B, natamycin, rimocidin, nystatin, and fludioxonil Lt; RTI ID = 0.0 &gt; 1, &lt; / RTI &gt; An antifungal agent comprising an inhibitor for an antifungal target transcription factor Hob1,
Wherein said inhibitor is an antibody to said protein. &Lt; Desc / Clms Page number 24 &gt; An antifungal agent comprising an inhibitor for a gene encoding an antifungal target transcription factor Hob1,
Wherein the inhibitor is an antisense oligonucleotide, siRNA, shRNA, miRNA, or a vector comprising the same, for the gene. The antifungal agent preparation for preventing, treating, or preventing and treating fungal infections of crops or animals. The antifungal agent preparation according to claim 24 or 25, which is administered in combination with an azole-based or non-azole-based antifungal agent, for preventing, treating or preventing and treating fungal infections in crops or animals. 27. The method of claim 26, wherein the azole-based antifungal agent is at least one selected from the group consisting of fluconazole, itraconazole, voriconazole, and ketoconazole. , An antifungal agent for the treatment or prophylaxis and treatment. 26. The method of claim 26, wherein the non-azole based antifungal agent is selected from the group consisting of amphotericin B, natamycin, rimocidin, nystatin, and fludioxonil Antifungal agent for preventing, treating or preventing and treating fungal infections of crops or animals. The antifungal agent screening method according to claim 1 or 2, wherein the cell is a fungal cell. 30. The method of screening an antifungal agent according to claim 29, wherein the fungus is bacillus or acacia. The antifungal agent screening method according to claim 1 or 2, wherein the antifungal agent is for preventing, treating, or preventing and treating fungal infections caused by basidiomycetes, acanthosis fungi, or acaricid bacteria and basidiomycetes. The antifungal agent screening method according to claim 1 or 2, wherein the antifungal agent is for preventing, treating, or preventing and treating a crop infection or an animal infection caused by basidiomycetes, acanthosis bacteria, or acaraceae bacteria. 32. The method of claim 31, wherein said bacillus or acarid is selected from the group consisting of Candida sp. , Aspergillus sp. , Cryptococcus sp. , Malassezia sp. Wherein the fungus is one or more fungi selected from the group consisting of Fusarium sp. And Penicillium sp. Fungi. 33. The method of claim 32, wherein said bacillus or acarid is selected from the group consisting of Candida sp. , Aspergillus sp. , Cryptococcus sp. , Malassezia sp. Wherein the fungus is one or more fungi selected from the group consisting of Fusarium sp. And Penicillium sp. Fungi.

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