Random Amplified Polymorphic DNA (RAPD) 분석법을 이용한 Vibrio vulnificus의 분류

조상원,김수현,신명근,신종희,서순팔,양동욱,조상원,김수현,신명근 대한임상병리학회 1997 대한임상병리학회지 Vol.17 No.6

장미속 유전자원의 유전적 관계와 국내 해당화의 유전적 다양성

정연화(Youn-Hwa Joung),김성태(Seung-Tae Kim),김기준(Gi-Jun Kim),이자현(Ja-Hyun Lee),기광연(Gwang-Yeon Gi),한태호(Tae-Ho Han) 한국원예학회 2010 원예과학기술지 Vol.28 No.6

Fifty-nine accessions of 23 species in genus Rosa were collected, and 15 accessions of Rosa rugosa were collected throughout 10 regions of Korea. Their genetic relationship was investigated by using morphological analysis and RAPD marker. The morphological analysis was measured for 7 quantitative and quantified for 4 qualitative traits. RAPD analysis obtained a total of 959 polymorphic bands by using twenty primers. Morphological analysis classified most according to the rose section system except for several species. The cluster analysis of genus Rosa based on RAPD data could identify the subgenus Platyrhodon and Eurosa. The subgenus Eurosa separated five sections; Gallicanae, Cinnamomeae, Pimpinellifolia, Synstylae, and Caninae. Correlation analysis between morphological and RAPD analysis showed low significance (r = 0.35). The accessions of R. rugosa belonged to the section Cinnamomeae clustered into three groups at genetic distance ranging from 0.28 on the base of RAPD analysis. In conclusion, the genetic relationship of the genus Rosa was consistent to the previously reported rose section system, and domestic collections of R. rugosa were separated from 3 groups on the base of RAPD marker.

Phylogenetic Study on the Genus Pungitius (Pisces: Gasterosteidae) Populations by the RAPD analysis통 연구

Chae, Byung-Soo,Seo, Bo-Keun 한국어류학회 2003 韓國魚類學會誌 Vol.15 No.1

한국산 가시고기속(genus Pungitius) 어류의 분류학적 위치를 밝히기 위하여 RAPD(random amplified polymorphic DNAs) method를 이용하여 분석하였다. 가시고기속 어류 중 P. kaibarae와 P. sinensis의 분류학적 위치에 대해서는 학자들 간에 이견이 많지만, 이들 2종은 뚜렷한 형태적 차이와 지리적 분포 양상에 의해 P. sinensis 집단 및 P. kaibarae 내의 3개 집단으로 구분된다. 집단간의 이러한 형태의 차이는 본 조사의 RAPD 분석 및 기존의 isoenzyme 분석에 의해 밝혀진 유전적 차이와 일치하였다. P. sinensis 집단과 P.kaibarae 집단 사이의 유전적 차이는 D = 0.772였으며, P. kaibarae 집단 내의 강릉 이북 집단과 금호강-형산강 집단 사이의 유전적 차이는 D = 0.680이었다. 이들 두 유전적 차이의 값은 별종 혹은 그 이상 수준의 차이에 해당하였다. 이들 두 유전적 차이의 값은 별종 혹은 그 이상 수준의 차이에 해당하였다. 한편 금호강과 형신강의 집단 사이의 유전적 차이는 D = 0.370이었으며 이는 아종수준의 차이에 해당하였다. 따라서 저자들은 유전적 및 형태적 차이를 근거로 P. sinensis 집단, P.kaibarae 집단 중 강릉 이북 집단 및 금호강-형산강 집단을 각기 별종으로 하고, 금호강 집단과 형산강 집단은 서로 아종으로 분류하였다. RAPD 분석은 기존의 방법보다 단순하고 쉬운 technique이며 계통 연구에 있어서 적절한 방법으로 판단되었다. Taxonomic relationships of the fishes in the genus Pungitius from Korea were analyzed using RAPD (random amplified polymorphic DNAs) method. The taxonomic status of P. sinensis and P. kaibarae have been disputed among researchers, but these two species can be divided into a P. sinensis group and three groups within P. kaibarae by their distinct morphological differences and geographical distribution patterns. The morphological differences among the groups accord with the genetic differences as revealed by RAPD analysis in the present study and by isoenzyme analysis in previous reports. The genetic difference between the P. sinensis and P. kaibarae groups was D = 0.772, and that between P. kaibarae groups of Sacheon river and Geumho-Hyeongsan river was D = 0.680. These two values correspond to species level or above. The genetic difference between Geumho and Hyeongsan river groups was D = 0.370, which means that the two groups are distinguishable at the subspecies level. On the basis of genetic and morphological differences, we classify the P. sinensis group, the northern group of P. kaibarae above Gangneung, and the Geumho-Hyeongsan river group as distinct species respectively, and classify the Geumho river group and Hyeongsan river group as subspecies. RAPD analysis was more simple and an easier technique than previous methods, and proved to be a proper method for phylogenetic studies.

Codominant Expression in RAPD Analysis

Kim, Sung Man,Kim, Yong Chul,Choi, In Soo,Lee, Chung Yeol,Park, Hyun Chul,Skorupska, Halina T. 密陽産業大學校 農業技術開發硏究所 1997 農業技術開發硏究所報 Vol.1 No.1

APD 分析에서 codominant marker를 選拔, 利用한다면 細密化된 遺傳子 地圖作成에 있어서 아주 용이하게 사용될 수 있다. 본 硏究의 目的은 RAPD 分析에서 codominant marker의 發見을 說明하고자 함이었다. 164개의 選拔된 RAPD marker들 중 하나의 primer, X01은 band pattern에 있어서 典型的인 codominant형을 나타내었다. F2:3세대 後代系統들의 분리비 分析에서 3:1 이나 1:2:1의 比率을 보였으며, 遺傳子 地圖作成에서도 X01¹, X01², X01¹²모두 같은 자리에 位置하였다. 이상의 결과들은 X01¹, X01²marker들이 codominant expression을 나타냄을 立證하는 證據들이다. Identification and utilization of codominant RAPD marker would be very useful for the genetic map saturation. The purpose of this study was to demonstrate the identification of codominant expression in RAPD analysis. One RAPD primer, X01, gave codominant banding pattern. In the segregation analysis of 79 F2:3 progenies. each marker showed the expected segregation ratio of 3 : 1 or 1 : 2 : 1. In the mapping analysis, both markers X01¹and X01²were localized in the same position as the two banded polymorphic pattern X01¹²on linkage group LGC-15. Our results suggest that two markers X01¹and X01²are inherited in a codominant mode. Further study for the elucidation of the mechanism for the occurrence of codominant expression in RAPD analysis is required.

Differences by RAPD-PCR Analysis within and between Rockfish (Sebastes schlegeli) Populations from the Yellow Sea and the Southern Sea in Korea

Yoon, Jong-Man,Kim, Jong-feon The Korean Society of Animal Reproduction 2001 Reproductive & developmental biology Vol.25 No.4

Polymerase chain reaction (PCR) amplification of DNA as 30 different arbitrary primers and random amplified polymorphic DNAs (RAPD) analysis were performed on genomic DNA extracted from the blood of the marine rockfish (Sebastes schlegeli) from the Yellow Sea and the Southern Sea. The unique properties of the genomic DNA were used to investigate the features of the population dynamics and origins of the species. Out of 30 primers, seven generated 207 highly reproducible RAPD polymorphic products, producing approximately 2.7 polymorphic bands per primer. About 67.4% of total amplified products (307) were either polymorphic (207) to rockfish. The degree of similarity varied from 0.22 to 0.63 as calculated by bandsharing analysis. Also, the average level of bandsharing was 0.39$\pm$0.02 within the rockfish strains. The electrophoretic analysis of RAPD-PCR products showed the relatively high levels if variation between different individuals in rockfish from the Yellow Sea. However, the RAPD outlines obtained with DNA of different rockfish strains from the Yellow Sea and the Southern Sea in Korea were very similar. Also, a small number of polymorphic bands were identified. Even if further analyses or more rockfish populations are required, this result implies RAPD analysis reflects genetic differences between the geographical strains of the rockfish.

RAPD分析法에 의한 참나무 優良種의 PCR反應 分析

趙顯瑞,李喆浩,李正煥 진주산업대학교 1999 산업과학기술연구소보 Vol.- No.6

RAPD를 이용한 참나무류의 band pattern은 primer의 종류에 따라 공시 수종별로 구분이 가능한 표식인자(marker)를 가지고 있었고, 상수리나무 (lane 1-11) band pattern에서는 고유 band를 보였다. Primer OPA-10, OPA-16에서 갈졸참나무 (lane 28)은 갈참나무(lane 22-25) 및 졸참나무 (lane 12-17)와 공유 band가 있었으마, 갈참나무와 유사한 band pattern을 보여 갈참나무와 유사한 경향을 보였다. 촐참나무 (lane 12∼17), 갈참나무 (lane23∼27)와 갈졸참나무(lane28) 집단 간에는 유사한 band pattern을 보여 갈졸참나무가 갈참나무에 근접한 교잡종으로 인식되었다. 군집분석결과 28개 집단은 7개 Grpup로 분류되었는데, 제1 집단은 상수리나무, 제 2 집단은 갈참나무, 제 3집단은 졸참나무 등으로 집단간 종의 구분이 가능하였다. In the Polymerase Chain Reaction (PCR) - Random Amplified Polymerase (RAPD) DNA analysis was performed on DNA samples obtained from leaf collected from wild Quercus spp. in Kyeongnam Province. PCR - DNA markers were used as a predictor of superior tree in future Quercus spp. breeding development. Eight primers were used for PCR and generated a total of 122 RAPD markers. According to the PCR products bands of Q. acutissima (lane 1-11), Q. serrata (lane 12-17), Q. uariabilis (lane 18-22), Q. aliena (lane 23-27) a n d Quercus x urticaefolia (lane28) were detected in all samples. Genetic relationships among the 28 individual samples were analyzed by the cluster analysis of 122 RAPD markers, which divides seven groups. According to the cluster analyses, First group are contains Q. acrtissima. Second group are Q. aliena, third group Q. serrata and so on. And close relationship between Quercus x urticaefolia and Q. aliena were ascertained by cluster analysis.

Genetic Linkage Mapping of RAPD Markers Segregating in Korean Ogol Chicken - White Leghorn Backcross Population

Hwang, K.C.,Song, K.D.,Kim, T.H.,Jeong, D.K.,Sohn, S.H.,Lillehoj, H.S.,Han, J.Y. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.3

This study was carried out to construct mapping population and to evaluate the methods involved, including polymorphic DNA marker system and appropriate statistical analysis. As an initial step to establish chicken genome mapping project, White Leghorn (WL) and Korean Ogol chicken (KOC) were used for generating backcross population. From 8 initial parents, total 280 backcross progenies were obtained and 40 were used for genotyping and linkage analysis. For development of novel polymorphic markers for KOC, Random Amplified Polymorphic DNA (RAPD) markers specific for this chicken line were generated. Also included in this study were six microsatellite markers from East Lansing map as reference loci. For segregation analysis, 15 RAPD markers and 6 microsatellites were used to genotype the backcross population. Among the RAPD markers that we developed, 2 pairs of markers were identified to be linked and another 4 RAPD markers showed linkage with microsatellites of known map. In summary, this study showed that our backcross population generated from the mating of KOC to WL serves as a valuable genetic resource for genotyping. Furthermore, RAPD markers are proved to be valuable in linkage mapping analysis.

금강제비꽃(Viola diamantiaca Nakai) 자생지의 환경특성과 RAPD 분석

서원복,유기억 한국식물분류학회 2011 식물 분류학회지 Vol.41 No.1

This study investigated the environmental factors and conducted a RAPD analysis for a better understanding of the environmental characteristics and regional genetic variation in samples from 18 different areas of Viola diamantiaca. The habitats are mostly located on the slopes of mountains facing north at an altitude ranging from 614 m to 1,462 m above sea level with angles of inclination ranging from 3 degrees to 30 degrees. A total of 268 vascular plant taxa are identified in 35 quadrates of 18 habitats. The importance value of V. diamantiaca is 11.58%, and four highly ranked species, Sasa borealis (5.61%), Meehania urticifolia (5.21%), Ainsliaea acerifolia (3.62%), Pseudostellaria palibiniana (3.60%) are considered to have an affinity with V. diamantiaca in their habitats. The degree of their average species diversity is 1.36, while this metric for their evenness and dominance are 0.89 and 0.07, respectively. The average field capacity of the soil is 25.99%, with organic matter at 17.47%, and the pH is 5.19. The soil texture was confirmed as sandy loam of eleven and loam of seven. The result of the RAPD analysis, among 78 bands amplified with a primer, 64 (84.6%) showed polymorphism. Eighteen populations could be classified into five groups with similarity coefficient values ranging from 0.53 to 0.86. The Mt. Jiri population, which is geographically segregated, shows basal branching within the 18 populations. Five populations, including two in the southern district in Gangwon-do and three in Chungcheongbuk-do, form a distinct clade. Four populations in the central district of Gangwon-do and Mt. Bohyeon in the Gyeongsangbuk-do clade form a sister to the clade containing two populations in Gyeonggi-do and five populations of the northern district in Gangwon-do. The Mt. Gariwang population is placed between the southern district and the central district in the Gangwon-do clades. 금강제비꽃 보전을 위한 기초자료를 확보하기 위해 18개 지역을 대상으로 자생지 환경특성과 RAPD를 이용한 집단 간 유연관계 분석을 실시하였다. 자생지는 해발 614−1,462 m의 범위에 위치하였으며, 경사는 3−30o로 비교적 완만하였다. 식생조사 결과 조사된 방형구 내에 출현한 관속식물은 총 268분류군이었으며, 초본층은 금강제비꽃의 중요치가 11.58%로 가장 높았고, 다음으로는 조릿대(5.61%), 벌깨덩굴(5.21%), 단풍취(3.62%), 큰개별꽃(3.60%) 등의 순으로 나타나 이 종류들이 금강제비꽃과 친화도가 높은 것으로 생각된다. 종다양도는 평균 1.36, 균등도는 평균 0.89, 우점도는 평균 0.07로 나타나 비교적 균일한 식생구조를 갖는 것으로 나타났다. 토양 분석결과 포장용수량, 유기물함량 그리고 pH는 각각 평균 25.99%, 17.47%, 5.19로 조사되었으며, 토성은 11개 지역이 사양토, 7개 지역이 양토로 확인되었다. RAPD 분석을 통해 총 78개의 band가 확인되었으며, 그 중 polymorphic band는 64(84.6%)개로 지역 간 높은 유전변이를 보였다. 18개 집단은 유사도지수 0.53−0.86 범위 내에서 5개의 분계조를 형성하였다. 지리적으로 가장 격리된 지리산 집단이 가장 기부에 위치하였고, 강원도 남부의 2개 집단과 충청북도의 3개 집단도 함께 유집되었다. 또한 강원도 중부에 위치한 4개 지역과 경상북도의 보현산이 하나의 분계조를 형성하였으며, 경기도의 2개 지역과 강원도 북부의 5개 지역이한 개의 군을 이루었고, 가리왕산 집단은 강원도 남부와 중부집단의 분계조 사이에 위치하였다.

RAPD(Random Amplified Polymorphic DNA)법을 이용한 한약재의 판별 연구

김대원,김도균,안선경,조동욱 한국한의학연구원 1997 한국한의학연구원논문집 Vol.3 No.1

Conventionally, identification and classification methods of natural products include the morphological survey and assay of chemical disposition. Using these methods, however, is not satisfying for the precise identification of natural products because they are often variable in the compositions, genomic DNA analysis such as RAPD, RFLP and Amp-FLP can be adopted for this purpose. In this study, various ginsengs and bear gall bladder were tested for the development of genetic identification and classification method. Varieties of ginsengs such as, P. ginseng, P. quinquefolium, P. japonicus and P. notohinseng, were genetically analyzed by RAPD. Also, DNA isolated form Bear blood and gall bladder, Ursus thibetanus, Ursus americanus and Ursus arctors, were analyzed by the same method. The results demonstrated that the identification and classification of bear gall bladder and various ginsengs were possible by RAPD analysis. Therefore, this method was thought to be used as a additional method for the identification of other natural products.

핵형분석과 RAPD技法을 利用한 상사화(Lycoris)속의 類綠關係 : Lycoris의 핵형 및 RAPD 분석

조기정,차재영,김완주,박윤점,박종군 圓光大學校大學院 2000 論文集 Vol.24 No.-

Twelve species of Lycoris was selected for establishment of phylogenetic relationships by karyotype and random amplified polymorphic DNA (RAPD) analyses. Our results obtained from karyotype analysis were very similar to those reported previously. For RAPD analysis, we used 10 synthetic oligonucleotides as primers. A total of 110 distinct DNA fragments were obtained by polymerase chain reaction. Pair-wise comparisons of unique and shared amplification products were performed to generate Nei and Li's similarity coefficients. On the basis of the dendrogram constructed with the similarity coefficients, 12 Lycoris species were divided into three clusters. Our results also suggested that L, incanata have originated from a cross between a gamete of L, sprengeri having 22A and another gamete having 4M+3T+1m.