• 한국어병학회지
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• 제19권2호
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• pp.141-153
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• 2006

Genomic DNA samples isolated from two geographical crayfish (Cambaroides similis) populations in the inland of the Korean Peninsula, at Jeonju (Jeonju crayfish; JJC) and Jeongup (Jeongup crayfish; JUe), were PCR-amplified repeatedly. The six arbitrarily selected primers OPC-03, OPC-06, OPC-09, URP-02, URP07 and URP-09 generated the common, specific, and polymorphic fragments. The sizes of DNA fragments also varied widely, from 100 bp - 2,600 bp. Here, 521 fragments were identified in the JJC population, and 354 in the JUC population: 6 primers generated 60 specific fragments (60/521 fragment, 11.5%) in the JJC population, and 90 (90/354 fragments, 25.4%) in the JUC population. These primers produced 42 polymorphic fragments (8.1%) in the DC population, and 18 (5.1%) in the mc population. Especially these results demonstrate that the primers detected numerous specific fragments. Especially, the decamer primer OPC-06 generated inter-population-common DNA fragments, approximately 400 and 800 bp, respectively, in both the JJC and JUC populations. The universal primer URP-02 also generated inter-population-identical DNA fragments, approximately 350 bp and 600 bp, between the two geographical crayfish populations. Based on the average bandsharing values of all samples, the bandsharing value of individuals within the JJC population was much higher than in the JUC population. The bandsharing value between individuals no. 10 and no. 15 was 0.683, which was the highest between the two geographical populations. The dendrogram obtained by the six primers indicates two genetic clusters: cluster I (CRAYFISH 01 - CRAYFISH II), and cluster 2 (CRAYFISH 12 - CRAYFISH 22). The genetic distance between the two geographical populations ranged from 0.053 to 0.605. Ultimately, the longest genetic distance displaying significant molecular differences was found to exist between individuals in the two crayfish populations, between individuals CRAYFISH no. 02 of Jeonju and CRAYFTSH no. 15 of Jeongup (genetic distance = 0.605).

• 한국양식학회지
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• 제19권2호
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• pp.67-76
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• 2006

Genomic DNA isolated from two Porphyra species, P. tenera and P. dentate from Wando located on the southern coast of Korean peninsula was amplified by PCR reaction. The amplified products were separated by agarose gel electrophoresis (AGE) with decamer primer and stained with ethidium bromide. The eight arbitrarily selected primers OPA-04, OPA-06, OPB-01, OPB-08, OPB-10, OPB-11, OPB-14 and OPC-10 generated the shared loci, polymorphic, and specific loci. The size of DNA bands varies from 100 bp to 2,200 bp. The complexity of the banding patterns varies dramatically between the primers and two Porphyra species. A total of 528 loci observed were identified in P. tenera and 443 in P. dentata: 22 polymorphic loci (4.2%) in P. tenera and 30 (6.8%) in P. dentata. 154 shared loci observed, the average 19.3 per primer, were identified in P. tenera and 143 loci, the aver-age 17.9 per primer, in P. dentata species. The number of specific loci in P. tenera and P. dentata was 73 and 77, respectively. The average bandsharing value was $0.623{\pm}0.008$ with P. tenera and $0.560{\pm}0.009$ within P. dentata. The average bandsharing value between two Porphyra species was $0.408{\pm}0.004$, ranged from 0.305 to 0.564. The dendrogram obtained by the eight primers indicates four genetic clusters. The genetic distance between two Porphyra species ranged from 0.076 to 0.627. The individual no. 02 of P. tenera was genetically closely related to no. 01 of P. tenera(genetic distance=0.082). Especially, two entities between the individual DENTATA no.21 and DENTATA no. 19 of P. dentata showed the longest genetic distance (0.627) in comparison with other individuals used. In this study, RAPD-PCR analysis has revealed the significant genetic distance between two Porphyra species pairs (P<0.001).

• 한국발생생물학회지:발생과생식
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• 제20권4호
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• pp.379-385
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• 2016

Seven oligonucleotides primers were shown to generate the shared loci, specific loci, unique shared loci to each species and shared loci by the three species which could be obviously scored. In the present study, 7 oligonucleotides primers produced 401 total loci in the Styela clava (SC) species, 390 in the Halocynthia roretzi (HR) and 434 in the Styela plicata (SP), respectively. Seven oligonucleotides primers generated 275 specific loci in the SC, 341 in the HR and 364 in the SP species, respectively. The oligonucleotides primer BION-23 generated 28 unique loci to each species in the SP species. Especially, the oligonucleotides primer BION-25 produced 7 unique loci to each species, which were identifying each species in the SP species. BION-17 distinguished 21 shared loci by the three ascidian species, major and/or minor fragments of sizes, which were identical in almost all of the samples. Based on the average bandsharing values of all samples, the similarity matrix ranged from 0.519 to 0.774 in the SC species, from 0.261 to 0.683 in the HR species and from 0.346 to 0.730 in the SP species. As regards average bandsharing value (BS) results, individuals from SC species ($0.661{\pm}0.081$) exhibited higher bandsharing values than did individuals from HR species ($0.555{\pm}0.074$) (P<0.05). The dendrogram obtained by the seven oligonucleotides primers indicates three genetic groups. In three ascidian species, the shortest genetic distance (0.071) exhibiting significant molecular difference was also between individual no. 20 and no. 21 within the SP species.

• 한국발생생물학회지:발생과생식
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• 제5권2호
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• pp.151-158
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• 2001

군산지역에 있는 호수와 양식장에서 채집된 붕어(Carassius carassius)의 혈액으로부터 추출된 genomic DNA를 무작위 primer를 이용한 PCR-RAPD 방법에 의해서 유전적 차이를 확인하고자 하였다. 12개 primer 중에서 6개를 이용한 결과 호수산 붕어의 경우 primer 당 약 2.1 polymorphic bands가 나타났고, 총 266개의 높은 RAPD marker가 확인되었으며, 0.18에서 0.76의 bandsharing분석 결과가 나타났다. 군산지역에 있는 호수와 양식장에서 채취된 붕어 2집단간의 RAPD 특징을 bandsharing value로 비교 분석해 본 결과 각각 호수산이 0.47, 양식산이 0.70 으로 나타났으며, 이는 양식산 개체들간에 유사성이 높게 나타났다. 이러한 결과는 군산지역에 있는 양식장의 경우 유사한 환경조건내에서 붕어가 사육되었거나 혹은 오랜 기간동안 근친교배의 결과 이러한 유전적 유사성이 높게 나타난 것으로 사료된다. 달리 말하면 비록 다양한 지리적인 분포가 있더라도 군산지역의 다른 지역으로부터 야생산 붕어 집단의 도입으로 인하여 genomic DNA의 높은 수준의 다양성을 가질 수 있다는 것이다. 일반적으로 primer에 의해서 제시된 RAPD 다형성은 양식대상 어종이면서 온수성 어종인 붕어의 계통 혹은 집단을 확인하기 위한 유전적 표지인자로서 사용될 수 있을 것이다. 그러나 앞으로 집단 및 채집장소의 추가적인 확보 그리고 다른 방법을 통한 연구가 미비한 점을 보완할 수 있는 데 필요하다고 사료된다.

• 한국발생생물학회지:발생과생식
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• 제16권4호
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• pp.315-320
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• 2012

The gDNA isolated from Cyclina sinensis from Gochang (GOCHANG), Incheon (INCHEON) and a Chinese site (CHINESE), were amplified by PCR. Here, the seven oligonucleotide decamer primers (BION-66, BION-68, BION-72, BION-73, BION-74, BION-76, and BION-80) were used to generate the unique shared loci to each population and shared loci by the three cyclina clam populations. As regards multiple comparisons of average bandsharing value results, cyclina clam population from Chinese (0.763) exhibited higher bandsharing values than did clam from Incheon (0.681). In this study, the dendrogram obtained by the seven decamer primers indicates three genetic clusters: cluster 1 (GOCHANG 01~GOCHANG 07), cluster 2 (INCHEON 08~INCHEON 14), cluster 3 (CHINESE 15~CHINESE 21). The shortest genetic distance that displayed significant molecular differences was between individuals 15 and 17 from the Chinese cyclina clam (0.049), while the longest genetic distance among the twenty-one cyclina clams that displayed significant molecular differences was between individuals GOCHANG no. 03 and INCHEON no. 12 (0.575). Individuals of Incheon cyclina clam population was somewhat closely related to that of Chinese cyclina clam population. In conclusion, our PCR analysis revealed a significant genetic distance among the three cyclina clam populations.

• 한국발생생물학회지:발생과생식
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• 제22권3호
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• pp.283-288
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• 2018

The author carried out PCR-based genetic platform to investigate the hierarchical polar dendrogram of Euclidean genetic distances of one bastard halibut population, particularly for Paralichthys olivaceus, which was further connected with those of the other fish population, by involving with the precisely designed oligonucleotide primer sets. Eight oligonucleotides primers were used generating excessively alterating fragments, ranging in size of DNA bands from larger than approximately 100 bp to less than 2,000 bp. As regards average bandsharing value (BS) results, individuals from Hampyeong population (0.810) displayed lower bandsharing values than did individuals from Wando population (0.877). The genetic distance between individuals approved the existence of close relationship in the cluster II. Relatively, individuals of one bastard halibut population were fairly related to that of the other fish population, as shown in the polar hierarchical dendrogram of Euclidean genetic distances. The points of a noteworthy genetic distance between two P. olivaceus populations demonstrated this PCR procedure is one of the quite a few means for individuals and/or populations biological DNA investigates, for species security and proliferation of bastard halibut individuals in coastal region of the Korea.

• 한국발생생물학회지:발생과생식
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• 제22권4호
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• pp.393-402
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• 2018

Genomic DNA was isolated from the Gracilaria vermiculophylla (GRV) and G. chorda (GRC) from Jangheung located in the southern sea of the Korean Peninsula, respectively and we performed clustering analyses, DNA polymorphisms and the genetic differences. The seven selected primers OPC-01, OPA-04, OPA-05, OPD-07, OPD-08, OPB-10, and OPD-16 generated average bandsharing (BS) value, the genetic distance and dendrogram. The size of DNA bands varies from 90 bp to 2,400 bp. The average BS value was $0.859{\pm}0.004$ within GRV and $0.916{\pm}0.006$ within GRC. The average BS value between two Gracilaria species was $0.340{\pm}0.003$, ranged from 0.250 to 0.415. The dendrogram obtained by the seven primers, indicates two genetic clusters. The genetic distance between two Gracilaria species ranged from 0.059 to 0.513. The individual VERMICULOPHYLLA no. 07 of GRV was genetically closely related to VERMICULOPHYLLA no. 06 of GRV (genetic distance=0.059). Especially, two entities between the individual VERMICULOPHYLLA no. 10 of GRV and CHORDA no. 22 of GRC showed the longest genetic distance (0.513) in comparison with other individuals used. Accordingly, as mentioned above, PCR analysis showed that the GRV was a little more genetically diverse than the GRC species. We convinced that this DNA analysis revealed a significant genetic distance between two Gracilaria species pairs (p<0.01).

• 한국어류학회지
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• 제17권3호
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• pp.173-186
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• 2005

한국과 대서양산 갈치 (Trichiurus lepturus) 2지리적 집단으로부터 genomic DNA를 분리 추출하였다. 선택된 8개의 RAPD primer를 이용하여 common, polymorphic 그리고 specific fragment를 얻어냈다. 2지역으로부터 primer간 banding patterns 의 복잡성이 두드러지게 나타났다. DNA fragment 의 분자적 크기는 150 bp에서부터 3,000 bp까지 커다란 차이를 나타내었다. 본 연구에서 한국산 갈치 집단에서는 947개의 fragment가 나타났고, 대서양산 갈치 집단에서는 642개의 fragment 가 확인되었다. 또한 한국산 집단에서는 148개의 specific fragment (15.6%) 가 확인되었으며, 대서양산 갈치집단에서는 61개의 specific fragment (9.5%)가 발생되었다. 한국산 갈치집단에서는 638개의 common fragment가 나타났으며, 이는 primer 당 평균적으로 79.8개의 fragment 로 확인되었다. 또한 대서양산 갈치집단에서는 429개의 common fragment 가 확인되었고, 평균해서 primer 당 53.6개의 common fragment 가 나타났다. 한국산 갈치집단과 대서양산 갈치집단의 polymorphic fragment는 각각 76개와 27개로 확인되었다. 모든 갈치시료의 평균적인 bandsharing value를 기초로 해서 한국산 갈치집단의 similarity matrix를 조사해 본 결과 0.784 로부터 0.922까지 나타났고, 대서양산 갈치집단의 값은 0.833로부터 0.990까지 확인되었다. 결과적으로 대서양산 갈치집단내 개체의 bandsharing value 평균값은 한국산 갈치집단의 평균값보다 높게 나타났다. 8개의 primer를 사용하여 얻어진 dendrogram은 cluster 1 (KOREAN 01~KOREAN 11) 및cluster 2 (ATLANTIC 12~ATLANTIC 22)와 같이 2개의 유전적 클러스터로 나뉘어졌다. 한국산 갈치집단내의 10번째 개체(KOREAN no. 10)와 11번째 개체(KOREAN no. 11) 사이가 가장 가까운 유전적 관계(genetic distance = 0.038)를 나타내었다. 궁극적으로 볼 때 한국산 갈치집단의 1번째(KOREAN no. 01)와 대서양산 갈치집단의 16번째(ATLANTIC no. 16) 개체 사이가 가장 먼 유전적 거리(genetic distance = 0.708)를 나타내었다.

• 한국양식학회지
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• 제17권1호
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• pp.69-80
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• 2004

The objective of the present study was to analyze genetic distances, variation and characteristics of individuals in rainbow trout, Oncorhynchus mykis using amplified fragment length polymorphism (AFLP) method as molecular genetic technique, to detect AFLP band patterns as genetic markers, and to compare the efficiency of agarosegel electrophoresis (AGE) and polyacrylamide gel electrophoresis (PAGE), respectively. Using 9 primer combinations, a total of 141 AFLP bands were produced, 108 bands (82.4%) of which were polymorphic in AGE. In PAGE, a total of 288 bands were detected, and 220 bands (76.4%) were polymorphic. The AFLP fingerprints of AGE were different from those of PAGE. Separation of the fragments with low molecular weight and genetic polymorphisms revealed a distinct pattern in the two gel systems. In the present study, the average bandsharing values of the individuals between two populations apart from the geographic sites in Kangwon-do ranged from 0.084 to 0.738 of AGE and PAGE. The bandsharing values between individuals No.9 and No. 10 showed the highest level within population, whereas the bandsharing values between individuals No.5 and No.7 showed the lowest level. As calculated by bandsharing analysis, an average of genetic difference (mean$\pm$SD) of individuals was approximately 0.590$\pm$0.125 in this population. In AGE, the single linkage dendrogram resulted from two primers (M11+H11 and M13+H11), indicating six genetic groupings composed of group 1 (No.9 and 10), group 2 (No. 1, 4, 5, 7, 10, 11, 16 and 17), group 3 (No. 2, 3, 6, 8, 12, 15 and 16), group 4 (No.9, 14 and 17), group 5 (No. 13, 19, 20 and 21) and group 6 (No. 23). In AGE, the genetic distances among individuals of between-population ranged from 0.108 to 0.392. In AGE, the shortest genetic distance (0.108) displaying significant molecular differences was between individuals No.9 and No. 10. Especially, the genetic distance between individuals No. 23 and the remnants among individuals within population was highest (0.392). Additionally, in the cluster analysis using the PAGE data, the single linkage dendrogram resulted from two primers (M12+H13 and M11+H13), indicating seven genetic groupings composed of group 1 (No. 15), group 2 (No. 14), group 3 (No. 11 and 12), group 4 (No.5, 6, 7, 8, 10 and 13), group 5 (No.1, 2, 3 and 4), group 6 (No.9) and group 7 (No. 16). By comparison with the individuals in PAGE, genetic distance between No. 10 and No. 7 showed the shortest value (0.071), also between No. 16 and No. 14 showed the highest value (0.242). As with the PAGE analysis, genetic differences were certainly apparent with 13 of 16 individuals showing greater than 80% AFLP-based similarity to their closest neighbor. The three individuals (No. 14, No. 15 and No. 16) of rainbow trout between two populations apart from the geographic sites in Kangwon-do formed distinct genetic distances as compared with other individuals. These results indicated that AFLP markers of this fish could be used as genetic information such as species identification, genetic relationship or analysis of genome structure, and selection aids for genetic improvement of economically important traits in fish species.

• 한국가축번식학회지
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• 제23권4호
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• pp.303-311
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• 1999

본 연구는 정자세포로부터 분리된 genome 내 DNA를 PCR 기법으로 증폭시킨 후 random amplified polymorphic DNA(RAPD) 분석을 통해 무지개송어의 품종 내 유전적 특성과 변이성을 해석하고 품종의 특이 유전적 표지를 개발하기 위해서 수행되었다. 20 종류의 primer를 사용하여 RAPD 양상을 검색한 후 다형현상의 출현빈도와 band 수에 기초하여 이들 중 6개의 primer를 선정하여 이용하였다. 그 중에 4개의 primer는 17개의 RAPD marker를 나타내었고, 그중 primer 당 8개인 48개 (28%)의 band 가 다형성을 보여주었다. 6개의 primer 중 4개는 개체들 사이에 다형성을 나타내는 band를 나타내었다. Bandsharing 의 경우 연어와 비교될 만큼 무지개송어는 3개의 특이적인 DNA marker를 가지고 있었다 (2.3. 2.0 및 1.3kb). 같은 무작위 primer를 이용해서 나타난 개별적인 band는 단일 primer 가 무지개송어의 정자핵 DNA의 경우 적어도 3개의 독립적인 genome 내 다형성을 탐지해 낼 수 있다는 것을 제시하고 있다. 이러한 primer에 의해서 나타난 RAPD 다형성은 개체식별을 위한 유전적 표지인자로서 사용될 수 있는 가능성을 제시하였으며, RAPD-PCR은 많은 어종에서 다형현상을 밝혀내는 기술이라 할 수 있다. 본 연구는 RAPD marker가 풍부하고 재현성이 있으며 RAPD 다형성을 지닌 marker를 사용하여 이러한 중요한 양식대상어종에서 미래의 gene mapping과 MAS 를 위한 기초를 제공해 줄 수 있다. RAPD 다형성은 어류의 품종 분화를 위한 유전적 표지로서 유용한 것으로 결론지을 수 있을 것이다.