KR100769358B1 - Method for Genetically Discriminating a Korean Native Porphyra yezoensis Ueda and a Primer Set for the Method - Google Patents
Method for Genetically Discriminating a Korean Native Porphyra yezoensis Ueda and a Primer Set for the Method Download PDFInfo
- Publication number
- KR100769358B1 KR100769358B1 KR1020060038989A KR20060038989A KR100769358B1 KR 100769358 B1 KR100769358 B1 KR 100769358B1 KR 1020060038989 A KR1020060038989 A KR 1020060038989A KR 20060038989 A KR20060038989 A KR 20060038989A KR 100769358 B1 KR100769358 B1 KR 100769358B1
- Authority
- KR
- South Korea
- Prior art keywords
- native
- laver
- base
- intron
- sequence
- Prior art date
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
도 1a 내지 1f는 일본산과 국내 자생품종 방사무늬김의 SSU rDNA의 상부 인트론의 염기서열을 정렬한 도표이다.Figures 1a to 1f is a table showing the alignment of the nucleotide sequence of the upper intron of SSU rDNA of domestic and domestic native varieties of radiation pattern laver.
도 2는 본 발명에 의한 PCR 증폭 기법에 따라 일본산과 자생품종의 SSU rDNA의 상부 인트론이 증폭됨을 보여주는 전기영동 사진이다.Figure 2 is an electrophoresis picture showing the amplification of the top intron of SSU rDNA of domestic and native varieties according to the PCR amplification technique according to the present invention.
삭제delete
본 발명은 방사무늬김 중에서 일본품종과 자생품종을 유전학적으로 간편하게 구별하기 위한 것으로, 보다 상세하게는 방사무늬김의 일본품종과 자생품종의 염기서열을 분석함으로써 유전학적으로 정확하면서도 간단하게 구별할 수 있는 방법에 관한 것이다.The present invention is to easily distinguish between the Japanese varieties and native varieties of the radiation pattern laver, more specifically, by analyzing the nucleotide sequence of the Japanese varieties and native species of the radiation pattern laver to be genetically accurate and simple to distinguish It is about how it can be.
우리나라 양식 김의 연간 생산량은 약 22.8만톤이고, 생산액은 1,915억원으로 전체 해조류 연간 총생산액인 2,560억원의 약 75%를 차지하고 있다(해양수산부 어업생산통계 2004).The annual output of aquaculture laver in Korea is about 22.8 million tons, and the output value is 155.1 billion yuan, accounting for about 75% of the total annual seaweed output of 260 billion yuan (Ministry of Maritime Affairs and Fisheries, 2004).
현재 국내에서는 참김(Porphyra tenera Kjellman), 방사무늬김(P. yezoensis Ueda), 모무늬돌김(P. seriata Kjellman) 그리고 잇바디돌김(P. dentata Kjellman) 등 4종이 주로 양식되고 있다. 인공채묘가 도입되기 전까지 우리나라에서 양식되어 온 김은 주로 참김으로 알려져 있다. 그러나 1968년부터 1970년까지 각 양식장의 표본을 채취하여 조사한 결과 참김은 일부 지역 또는 한 시기에 국한되어 양식되는 반면 방사무늬김은 대부분의 양식장에서 주류를 이루어 양식되는 것을 확인한 바 있다. 이러한 방사무늬김의 강세현상은 향후 인공채묘가 보급됨에 따라 더욱 가속화되었다(강제원, 한국수산학회지 3: 77-92, 1970). Currently, four species of seaweeds ( Porphyra tenera Kjellman), Radial patterned seaweed ( P. yezoensis Ueda), P. seriata Kjellman and P. dentata Kjellman are mainly farmed. Seaweed that has been farmed in Korea until artificial seedlings were introduced is known as sesame. However, from 1968 to 1970, sampling of the farms and surveys confirmed that sesame seeds were farmed in some regions or at one time, while radioactive laver was found in most farms. This strong pattern of radial laver was further accelerated by the spread of artificial seedlings in the future (Kang, Je-Won, Journal of the Korean Fisheries Society, 3: 77-92, 1970).
현재 방사무늬김은 우리나라 양식김의 약 70%를 차지하고, 김밥용 김(190×210 mm)과 재래김(190×270 mm)으로 가공되고 있으며, 품종으로는 일본에서 도입된 품종과 자생품종으로 구분된다. Radiation pattern laver accounts for about 70% of Korean laver and is processed into laver (190 × 210 mm) and traditional laver (190 × 270 mm). Are distinguished.
방사무늬김을 생산하고 있는 어업인들은 일본품종 또는 자생품종을 구분하여 공급해 줄 것을 종묘생산업계에 요구하고 있다. 그러나 김은 생육환경에 따라 형태·해부학적 형질 변이가 크기 때문에(황 등 Algae 16: 233-273, 2001) 이러한 형질만으로는 방사무늬김의 일본품종과 자생품종을 구분하기 어려우므로 종묘생산업체 에서는 품종을 명시하지 않은 상태로 종묘(사상체)를 판매하고 있으며 이로 인해 품종관리가 전혀 이루어지지 않고 있을 뿐 아니라 품종과 관련된 크고 작은 분쟁의 원인이 되고 있다. 따라서 김의 정상적인 품종관리 및 품질개선과 다양화를 위해 특정 염기서열과 primer 개발이 요구되고 있다. Fishermen who produce radiant laver are demanding the seedling production industry to supply Japanese or native varieties separately. However, because the morphological and anatomical trait variation is large depending on the growing environment (Hwang et al., Algae 16: 233-273, 2001), it is difficult to distinguish between Japanese and native varieties of radiant laver by these traits. Seedlings are sold in the absence of a stipulation, which causes not only varieties management but also causes large and small disputes related to varieties. Therefore, specific sequencing and primer development are required for normal varieties management, quality improvement and diversification of seaweed.
현재까지 김을 대량으로 생산하고 있는 국가는 한국, 일본, 중국이며, 한국과 중국에서는 방사무늬김의 경우 상당 부분 일본품종이 사상체 형태로 유통되고 있는 실정이다. 종자전쟁이라고도 불리는 신품종보호제도를 대비하는 차원에서 국내 자생품종을 정확히 식별하고 이에 육종기술을 추가하여 신품종을 개발한다면 일본품종에 대한 로열티 지급을 최대한 방지하고, 나아가 자생품종 유래의 신품종 수출을 통하여 로열티 수익을 창출할 수 있게 될 것이다.So far, the countries producing large amounts of laver are Korea, Japan, and China, and in Korea and China, a large part of Japanese varieties are distributed in the form of Sasang bodies. In preparation for a new breed protection system, also called seed war, if a native breed is correctly identified and breeding technology is developed, new breeds can be prevented and royalties can be prevented as much as possible through the export of new breeds derived from native breeds. You will be able to make money.
본 발명은 상기와 같은 종래기술의 문제점을 해결하기 위한 것으로, 염기서열 분석을 통해 보다 정확하면서도 간편한 품종 구별방법을 제공하고자 하는 것을 목적으로 한다.The present invention is to solve the problems of the prior art as described above, it is an object of the present invention to provide a more accurate and simple method for distinguishing varieties through sequencing.
전술한 목적을 달성하기 위한 본 발명은 핵의 SSU (small subunit) rDNA의 상부 인트론의 염기서열을 비교함으로써 방사무늬김 중에서 일본품종과 자생품종을 유전학적으로 간편하게 구별할 수 있는 방법을 제공한다.In order to achieve the above object, the present invention provides a method for genetically and easily distinguishing Japanese varieties and native varieties among radiant laver by comparing the nucleotide sequences of the upper introns of the small subunit (SSU) rDNA of the nucleus.
이를 위하여 본 발명자는 먼저 국내 각지의 자연 및 양식장에서 김 샘플을 다수 채집한 다음 종래 알려진 방법(예컨대, 황미숙 등 논문 "한국 남서해안 자연채묘 양식 김의 DNA 염기서열과 동정" Algae, 20(3); 183-196, 2005)에 따라 동정하여 방사무늬김 9개 시료를 선별하였다(표 2 참조).To this end, the present inventors first collect a large number of laver samples from nature and farms in Korea, and then known methods (e.g., Misook Hwang et al., "DNA Sequences and Identification of Natural Seed Cultured Laver on the Southwest Coast of Korea" Algae, 20 (3) 183-196, 2005), and nine samples of the patterned laver were selected (see Table 2).
이어서 선별된 각 방사무늬김의 총 DNA를 추출하고 증폭하여 SSU rDNA의 상부(upstream) 인트론(intron)의 염기서열을 분석하였다. 분석된 염기서열을 이미 알려진 일본산 방사무늬김의 동일 염기서열(서열1)과 정렬하여 비교함으로써 SSU의 상부 인트론 염기서열의 특정 부위가 상이함을 확인하였다(도 1a~1f 참조). 일본산 방사무늬김의 SSU rDNA 인트론의 염기서열은 GenBank에서 입수하였다. 선별된 시료 중에서 한 종류는 일본산의 염기서열과 일치하였는데, 이는 양식과정에서 일본산 방사무늬김이 혼입되었기 때문인 것으로 판단된다. Subsequently, the total DNA of each selected spine laver was extracted and amplified to analyze the nucleotide sequence of the upstream intron of the SSU rDNA. By comparing the analyzed nucleotide sequence with the same nucleotide sequence (SEQ ID NO. 1) of the known Japanese radiant pattern laver, it was confirmed that a specific site of the upper intron nucleotide sequence of the SSU is different (see FIGS. 1A to 1F). The nucleotide sequence of SSU rDNA intron of Japanese radish laver was obtained from GenBank. One of the selected samples was consistent with the Japanese nucleotide sequence, which is believed to be due to the incorporation of Japanese radish laver during the aquaculture process.
SSU rDNA의 상부 인트론의 염기서열을 결정하기 위하여 PCR용 프라이머 세트(서열2 및 서열3)를 개발하였다(청구항 10).Primer sets (SEQ ID NO: 2 and SEQ ID NO: 3) for PCR were developed to determine the nucleotide sequence of the upper intron of SSU rDNA (Claim 10).
이에 따라 본 발명은, (A) 방사무늬김 자생품종의 SSU rDNA의 상부(upstream) 인트론의 염기서열을 결정하고, 일본산 방사무늬김의 SSU rDNA의 상부 인트론의 염기서열(서열 1)과 정렬한 다음, (B) 정렬된 염기서열에서 12, 71, 75, 82, 251, 331, 348번째 염기 중 어느 하나 이상이 T이거나, 343, 495번째 염기가 A이거나, 353, 356, 496번째 염기 중 어느 하나 이상이 G이거나, 485, 486번째 염기 중 어느 하나 이상이 C이면 자생품종으로 판단하는 단계를 포함하는 방사무늬김 자생품종의 유전학적 구별방법을 제공한다(청구항 1).Accordingly, the present invention, (A) determines the nucleotide sequence of the upstream intron of SSU rDNA of native patterned radish laver, and aligns with the nucleotide sequence of the upper intron of SSU rDNA of Japanese radiant laver (SEQ ID NO: 1). (B) at least one of the 12, 71, 75, 82, 251, 331, and 348 bases in the aligned base sequence is T, the 343, 495 base is A, or the 353, 356, 496 bases. If any one or more of G, or if any one or more of the 485, 486th base is C provides a method of genetic distinction of the radioactive pattern of the native pattern including the step of determining the native species (Claim 1).
자생품종과 일본산 방사무늬김의 SSU rDNA 상부 인트론의 염기서열을 정렬하고 비교한 바, 도 1a 내지 도 1f에 나타난 바와 같이 14개소의 염기가 상이함을 확인할 수 있었다(이를 표 4에 정리하였다). 본 발명의 제 1항은 이들 14개소 염기 중에서 어느 하나라도 상이함이 확인되면 자생품종인 것으로 결정하는 방법이다.As a result of aligning and comparing the nucleotide sequences of the SSU rDNA upper introns of native varieties with Japanese radiant laver, as shown in FIGS. 1A to 1F, it was confirmed that 14 bases were different (Table 4 is summarized in Table 4). ).
한편, 전술한 바와 같은 염기서열 변이는 인접한 위치에서 3~5개소의 변이가 집중적으로 나타난다. 따라서 본 발명은 71, 75, 82번째 염기가 모두 T이거나(청구항 2), 331, 343, 348, 353, 356번째 염기가 각각 T, A, T, G, G 이거나(청구항 3), 485, 486, 495, 496번째 염기가 각각 C, C, A, G이면(청구항 4) 더욱 확실하게 자생품종으로 결정할 수 있게 된다.On the other hand, in the nucleotide sequence variation as described above, three to five variations appear intensively in adjacent positions. Thus, in the present invention, the 71st, 75th, 82nd bases are all T (claim 2), the 331, 343, 348, 353, 356th bases are T, A, T, G, G (claim 3), 485, If the 486, 495, and 496th bases are C, C, A, and G (claim 4), they can be more reliably determined as native species.
본 발명에 의한 방사무늬김 자생품종의 유전학적 구별방법을 수행하기 위하여, 방사무늬김의 총 DNA로부터 SSU rDNA의 상부 인트론을 PCR 증폭한 다음 염기서열을 결정하는 것이 바람직하다. 이러한 PCR 증폭을 위하여 InF1(서열2) 및 InR1(서열3)의 프라이머 세트를 사용한다(청구항 5). 이때 사용되는 상기 프라이머 세트에 관해서도 별도의 청구항으로 청구하였다(청구항 10). In order to carry out the genetic discrimination method of the native pattern of the radiation patterned kimchi according to the present invention, it is preferable to PCR amplify the upper intron of the SSU rDNA from the total DNA of the radiation patterned kimchi and then determine the base sequence. For this PCR amplification, primer sets of InF1 (SEQ ID NO: 2) and InR1 (SEQ ID NO: 3) are used (claim 5). The primer set used in this case was also claimed in a separate claim (claim 10).
삭제delete
삭제delete
본 발명에서 사용되는 프라이머 세트의 서열을 표 1에 정리하여 나타내었다.The sequences of the primer sets used in the present invention are shown in Table 1.
이하 실시·적용예를 통하여 본 발명을 상세하게 설명한다. 이들 실시·적용예는 본 발명의 확립과정과 적용예를 동시에 설명하기 위한 예시적인 것일 뿐 본 발명의 기술적 사상 또는 그 범위가 이에 의해 변하거나 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail through examples. These embodiments and application examples are merely illustrative for explaining the establishment process and application examples of the present invention, but the technical spirit or scope of the present invention is not changed or limited thereto.
실시·적용예 1 : 시료의 채집 및 방사무늬김의 선별Example 1 Application of Samples and Screening of Radiation Strips
우리나라의 서해안 및 남해안의 각지의 양식장 및 비양식장(자연 암반)에서 시료를 채집하였다. 시료는 멸균 해수용액으로 부착생물과 오염물질을 제거하고 Kimwipes로 수분을 제거한 후 건조기가 담긴 원심분리용 튜브에 넣어 상온에 보관하였다. 채집된 시료를 황 등의 방법("한국 남서해안 자연채묘 양식 김의 DNA 염기서열과 동정" Algae, 20(3); 183-196, 2005)에 따라 동정하여 방사무늬김 9개 시료를 선별하였다(표 2).Samples were collected from farms and non-farms (natural rock) in the west and south coasts of Korea. Samples were stored at room temperature by removing adherents and contaminants with sterile seawater, removing water with Kimwipes, and placing them in a centrifuge tube containing a dryer. The collected samples were identified according to the method of Hwang et al. ("DNA Sequence and Identification of Natural Seed Culture Kim, Southwest Coast of Korea" Algae, 20 (3); 183-196, 2005). (Table 2).
실시·적용예 2 : DNA 분리, 정제 및 SSU rDNA 염기서열 결정Example 2 Application: DNA Separation, Purification and SSU rDNA Sequence Determination
선별된 방사무늬김 시료로부터 총 DNA를 추출한 다음 이를 주형으로 하여각각의 SSU rDNA 염기서열을 분석하였다.Total DNA was extracted from the selected radiation patterned samples, and each SSU rDNA sequence was analyzed as a template.
(1) DNA 분리 정제 (1) DNA isolation and purification
건조 보관한 시료를 Strach-Crain 등(1997)의 방법으로 세포를 분해하고 단백질을 제거한 후 Wizard DNA Clean-Up System (Promega, WI, USA)의 resin을 1mL 첨가하여 column을 통과시켰다. 이어서 2㎕의 80% isopropanol을 column에 통과시킨 후 2분간 12,000rpm으로 원심분리한 다음 10분간 건조하였다. 이어서 50㎕의 증류수를 column에 넣고 20초간 12,000rpm으로 원심분리하여 DNA를 추출하였다. 추출한 DNA는 에티디움브로마이드(EtBr)가 첨가된 0.8% agarose gel 상에서 확인하였다.The dried samples were digested with Strach-Crain et al. (1997) method, protein was removed, and 1 mL of resin from Wizard DNA Clean-Up System (Promega, WI, USA) was added to the column. Subsequently, 2 μl of 80% isopropanol was passed through the column, followed by centrifugation at 12,000 rpm for 2 minutes, followed by drying for 10 minutes. Subsequently, 50 µl of distilled water was added to the column and centrifuged at 12,000 rpm for 20 seconds to extract DNA. The extracted DNA was identified on 0.8% agarose gel with ethidium bromide (EtBr).
(2) SSU rDNA 염기서열 분석 (2) SSU rDNA sequencing
추출된 총 DNA를 주형으로 하고, 사전에 확인된 일본산 방사무늬김의 핵 SSU rDNA 염기서열을 참조하여 제작한 InF1(서열2) 및 InR1(서열3) 프라이머 세트를 사용를 사용하여 각 시료에 대한 PCR을 수행하였다(도 2). 도 2 및 3에서 A: D976, B: NS06, C: HM012, D: HM014, E: NS01, F: NS02, G: NS03, H: NS04, I: NS05, J: NS07를 나타낸다.Using the extracted total DNA as a template, each sample was prepared using a set of InF1 (SEQ ID NO: 2) and InR1 (SEQ ID NO: 3) primers prepared by referring to the nuclear SSU rDNA nucleotide sequence of a Japanese radiation patterned kimchi previously identified. PCR was performed (FIG. 2). 2 and 3 show A: D976, B: NS06, C: HM012, D: HM014, E: NS01, F: NS02, G: NS03, H: NS04, I: NS05, J: NS07.
증폭된 유전자 부위의 PCR 산물을 DNA 정제 키트를 사용하여 정제한 다음 서열분석 키트(BigDyeTM Terminator Cycle Sequencing Ready Reaction Kit; ABI, USA) 및 DNA 분석기(Applied Biosystems 3730 DNA Analyzer; Applied Biosystems, USA)로 각 시료에 대한 염기서열을 결정하였다.The PCR product of the amplified gene region was purified using a DNA purification kit, followed by a sequencing kit (BigDye ™ Terminator Cycle Sequencing Ready Reaction Kit (ABI, USA)) and a DNA analyzer (Applied Biosystems 3730 DNA Analyzer; Applied Biosystems, USA). The base sequence for each sample was determined.
실시·적용예 3 : SSU rDNA 염기서열의 정렬 및 염기서열 변이 분석Example 3: Application of SSU rDNA Sequence and Alignment Sequence Analysis
분석된 각 시료 및 일본산 방사무늬김의 SSU rDNA의 상부 인트론의 염기서열을 Clustral X Program(v.1.64b, Thompson et al., 1997)을 사용하여 정렬시킨 후 MacClade Program(v.3.08a, Maddison and Maddison, 1999)을 사용하여 염기서열의 변화를 확인하였다(도 1a~도 1f). 도 1a~도 1f에서 P.ye는 방사무늬김인 P. yezoensis Ueda를 나타내며, 이어서 시료명을 기재하였다. 예를 들면, P.yeAB177은 시료명 AB177인 일본 홋카이도 하코다테의 자연산 방사무늬김을 의미한다.Sequences of each sample analyzed and the top intron of SSU rDNA of Japanese radiant laver were sorted using Clustral X Program (v.1.64b, Thompson et al., 1997), followed by MacClade Program (v.3.08a, Maddison and Maddison, 1999) was used to confirm the change in the nucleotide sequence (Fig. 1a to 1f). 1A to 1F, P.ye represents P. yezoensis Ueda, which is a radiation pattern, followed by a sample name. For example, P.yeAB177 means a natural radiant laver from Hakodate, Hokkaido, Japan, with sample name AB177.
비교를 위한 일본산 방사무늬김의 출처는 표 3과 같으며, 이들의 SSU rDNA 인트론의 염기서열은 GenBank에서 입수하였고 이들 중 D976(TU-1 culture)은 홋카이도대학에서 실물시료를 분양받아 실제 염기서열 분석에 활용하였다. Sources of Japanese radiant patterned seaweed for comparison are shown in Table 3.The base sequences of these SSU rDNA introns were obtained from GenBank, and among them, D976 (TU-1 culture) was obtained from real samples from Hokkaido University. It was used for sequence analysis.
분석된 각 시료와 일본산 방사무늬김의 염기서열 변이를 검토한 바, 시료 NS06을 제외한 모든 시료는 14개소에서 일본산과 다른 염기를 나타내었다(표 4). 김 양식장에서는 일본품종과 자생품종이 함께 양식되고 있으므로, 예외가 된 NS06은 일본품종으로 판명될 수 있다. As a result of examining the sequence variation of each sample analyzed and Japanese radiant laver, all of the samples except for the sample NS06 showed different bases from those produced in Japan (Table 4). In the seaweed farm, Japanese varieties and native varieties are grown together, so NS06, which is an exception, may turn out to be a Japanese variety.
따라서 본 발명에 의한 방법에 따라 방사무늬김의 SSU rDNA의 상부 인트론의 염기서열을 분석하고 검토하는 경우 간편하고 정확하게 일본산과 한국 고유의 자생품종을 구분할 수 있게 된다.Therefore, when analyzing and examining the nucleotide sequence of the upper intron of the SSU rDNA of the radiation patterned laver according to the method of the present invention, it is possible to easily and accurately distinguish between native and native Korean varieties.
삭제delete
삭제delete
삭제delete
삭제delete
삭제delete
삭제delete
삭제delete
본 발명에 의하면 방사무늬김의 품종관리 및 제품(김)의 품질관리를 용이하게 할 수 있고 나아가 본 발명의 프라이머 세트를 이용한 검출키트를 활용하여 비전문가에 의해서도 간편하게 품종관리 및 품질관리가 가능하게 된다.According to the present invention, it is possible to facilitate the varieties management of the radiation pattern laver and the quality control of the product (kim), and furthermore, by utilizing the detection kit using the primer set of the present invention, the varieties management and quality control can be easily performed even by non-experts. .
또한, 본 발명에 의해 방사무늬김의 자생품종을 정확히 동정하고 이에 육종기술을 추가하여 자생 신품종을 개발할 기반을 확립할 수 있다.In addition, according to the present invention, it is possible to accurately identify the native breed of the radiation pattern laver and add a breeding technique thereto to establish a foundation for developing a native native breed.
서열목록 전자파일 첨부 Attach sequence list electronic file
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020060038989A KR100769358B1 (en) | 2006-04-28 | 2006-04-28 | Method for Genetically Discriminating a Korean Native Porphyra yezoensis Ueda and a Primer Set for the Method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020060038989A KR100769358B1 (en) | 2006-04-28 | 2006-04-28 | Method for Genetically Discriminating a Korean Native Porphyra yezoensis Ueda and a Primer Set for the Method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR100769358B1 true KR100769358B1 (en) | 2007-11-01 |
Family
ID=39060362
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020060038989A KR100769358B1 (en) | 2006-04-28 | 2006-04-28 | Method for Genetically Discriminating a Korean Native Porphyra yezoensis Ueda and a Primer Set for the Method |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR100769358B1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101387056B1 (en) | 2012-03-12 | 2014-04-25 | 서천군 | Cultivating method of novel Porphyra yezoensis seopoong |
| KR101725316B1 (en) | 2016-06-24 | 2017-04-26 | 대한민국 | Method for identification of varieties in Pyropia yezoensis by using the structural variation of mitochondrial rnl gene |
| KR102290111B1 (en) | 2020-05-08 | 2021-08-17 | 전남대학교 산학협력단 | Primer Sets for Detecting SSR markers for Grouping of Pyropia yezoensis Cultivas, and Method for Grouping of Pyropia yezoensis Cultivas Using the Composition |
| KR20220153926A (en) | 2021-05-12 | 2022-11-21 | 전남대학교산학협력단 | SSR Marker for Identification of Blade type of Pyropia yezoensis and Application thereof |
-
2006
- 2006-04-28 KR KR1020060038989A patent/KR100769358B1/en active IP Right Grant
Non-Patent Citations (1)
| Title |
|---|
| 논문 |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101387056B1 (en) | 2012-03-12 | 2014-04-25 | 서천군 | Cultivating method of novel Porphyra yezoensis seopoong |
| KR101725316B1 (en) | 2016-06-24 | 2017-04-26 | 대한민국 | Method for identification of varieties in Pyropia yezoensis by using the structural variation of mitochondrial rnl gene |
| KR102290111B1 (en) | 2020-05-08 | 2021-08-17 | 전남대학교 산학협력단 | Primer Sets for Detecting SSR markers for Grouping of Pyropia yezoensis Cultivas, and Method for Grouping of Pyropia yezoensis Cultivas Using the Composition |
| KR20220153926A (en) | 2021-05-12 | 2022-11-21 | 전남대학교산학협력단 | SSR Marker for Identification of Blade type of Pyropia yezoensis and Application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108998547A (en) | A kind of microsatellite marking method for C. guichenoti paternity test | |
| CN105603098A (en) | Microsatellite marker primers used for penaeus monodon microsatellite family identification, identification method and application | |
| CN109517920B (en) | Construction method and application of fingerprint of eremochloa ophiuroides germplasm | |
| CN104004828B (en) | A kind of molecule marker and application thereof identifying onion cytoplasmic type | |
| KR100769358B1 (en) | Method for Genetically Discriminating a Korean Native Porphyra yezoensis Ueda and a Primer Set for the Method | |
| CN101418347A (en) | Yellow catfish sex chromosome specific molecular markers and genetic sex identification method | |
| CN111733274B (en) | SNP locus, primer set and method for identifying cauliflower Zhe nong piny flower 80-day variety | |
| CN105506162A (en) | SNP marker related to rapid growth of pacific oysters and identification method as well as application thereof | |
| CN112176074A (en) | Real-time fluorescent PCR primer probe and method for detecting patinopecten yessoensis | |
| CN105543222B (en) | The molecular labeling InDeL_33 of soybean 100-grain weight main effect QTL and its application | |
| CN103525814B (en) | A kind of special SCAR mark of Cynoglossus semilaevis sex and application method | |
| CN102876777B (en) | The special primer of brown croaker EST microsatellite marker and screening method | |
| CN101117645A (en) | Molecule labeling method for non-heading Chinese cabbage late bolting gene | |
| CN103911444B (en) | Primer for discriminating anguilla marmorata and anguilla bicolor pacifica fry and method thereof | |
| CN101974516B (en) | Method for discriminating Chinese milk vetch variety by using SSR (Simple Sequence Repeat)fingerprint spectrum | |
| CN108220456B (en) | EST-SSR primer and identification method for identifying crassostrea hongkongensis, crassostrea sikamea and first filial generation thereof | |
| CN102952797B (en) | SCAR marker closely linked with onion male sterility gene ms and application thereof | |
| CN109679967A (en) | One anti-soybean mosaic virus gene GmRDBMApplication | |
| CN102925431B (en) | SCAR marker closely linked to onion male sterile restoring gene Ms and application thereof | |
| CN115679004A (en) | Primers, method and kit for identifying Pelteobagrus vachelli, leiocassis longirostris and hybrid species | |
| CN111118130B (en) | Method for rapidly identifying sex of macrobrachium rosenbergii | |
| CN108359745A (en) | The dual RT-PCR method of synchronous detection wheat dwarf virus and Wheat Yellow strip virus | |
| CN102534039A (en) | Quick identification method and application of transgenic fish homozygote | |
| CN109022610B (en) | Molecular specificity marker primer of anoectochilus formosanus and identification method thereof | |
| CN102154287A (en) | Molecular marker for Fenneropenaeus chinensis with high-pH resistance character and identification method using same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant |