• 생명과학회지
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• 제11권1호
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• pp.65-67
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• 2001

In order to develop a rapid and simple method for testing the purity of radish hybrid seeds using a procedure based on the PCR(Polymerase chain reaction), eighty random primers were screened with the genomic DNA extracted from five day old seedlings of inbred parent lines and their F1 hybrids. Two primers, HRM-02 (5'-GAGACCAGAC-3') and HRM-19(5'-TGAGGCGTGT-3'), generate reproducible unique PCR patterns which can identify each parent lines as well as their hybrids. In actual test of randomly selected hybrid seeds using the two marker primers, the purity tested by one primer was exactly same as that of other primer. It suggests that one marker primer selected in this experiment is enough for the purity test of radish hybrid seeds. We demonstrates the use of RAPD(randomly amplified polymorphic DNAs) markers to identify each of inbred parent lines and hybrids by rapid and simple method.

• Plant Resources
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• 제4권3호
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• pp.121-129
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• 2001

Intraspecific genetic diversity of sixteen accessions of Mogolian Alliums including fifteen species was investigated using randomly amplified polymorphic DNA (RAPD) analysis. Twenty three out of forty primers revealed scorable polymorphism. A total of 440 RAPD markers were generated on the 16 accessions of Mongolian Alliums. Among 440 RAPDs assayed, 439 were polymorphic with a mean polymorphic rate of 99.7%. Unweighted pair-group method using an arithmetic average (UPGMA) cluster analysis using RAPD data separated the 16 Allium accessions into two broad groups at similarity index 0.70. The clustering of the species was closely related with previous classification between A. altaicum and A. fistulosum. In addition, a high genetic similarity was showed between A. cepa and A. tagar.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.85.1-85.1
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• 2003

Previously, we found the operon random primer (OP-5A) that is characteristic the genus Panax by randomly amplified polymorphic DNA (RAPD) analysis. However, OP-5A primer is limited to apply on the differentiation of only crude herbal plants. To construct more sensitive and unique primers on the genus Panax, ginseng-specific DNA profile (350 bp) that was amplified by OP-5A primer were inserted in a plasmid vector in the TA cloning method and sequenced. We designed the PCR primers (Forward: 5＂-AGGGGTCTTGCTAT AGCGGAAC-3＂, Reverse: 5＂-AGTCTTAATTTCATATTTTCGTATG-3＂) and identified the unique ginseng band (350 bp) in commercial granule products including ginseng extracts as well as crude ginseng plants by nascent PCR.(omitted)

• 한국약용작물학회지
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• 제4권3호
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• pp.224-230
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• 1996

서철재래종, 내장산 수집종, 대둔산 수집종, 거제도 수집종, 진부재래종 및 일본 도입종 등 계통의 용담을 대상으로 RAPD 분석을 통한 유사도 분석을 수행한 결과 적용한 20가지의 10-merprimer 중 8가지에서 54개의 증폭된 DNA 절편을 얻었다. 그 중 53.7%에 해당하는 29개의 band가 다형현상을 보였으며, 9.3%에 해당하는 5개의 band는 모든 계통에서 공통적으로 나타났고, 37%인 20개의 band가 특이성을 보였다. 특이성을 보이는 band들은 내장산 수집종에서 2개, 거제도 수집종에서 1개, 진부재래종에서 11개, 일본 도입종에서 6개가 관찰되었다. 유전적 유사도의 분석결과 서천재래종, 내장산 수집종, 대둔산 수집종 및 거제도 수집종이 0.76-0.87의 유사도를 보여 한 집단으로 구분되었으며, 이 집단과 진부재래종 및 일본 도입종 사이의 유사도는 0.23-0.34로 각 각 다른 집단으로 구별되었다. 또한 진부재래 종과 일본 도입 종 사이의 유사도는 0.41로 나타나 서로 구분되었다.

• Journal of Plant Biology
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• 제39권3호
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• pp.185-188
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• 1996

Randomly amplified polymorphic DNAs were employed to study the genetic variation and gene introgression in potato dihaploids (2n=24) which were generated after interspecific pollination of tetraploid cultivars (2n=4X=48, Solanum tuberosum cv Irish Cobbler, Superior and Dejima) by haploid inducer clones (2n=2X=24, Solanum phureja 1.22, Hes-5 and Hes-6). Genetic variation and DNA marker segregation among dihaploids were observed. Most dihaploids contain S. tuberosum specific RAPD markers but haploid inducer-specific RAPD markers were also found in some dihaploids. Of six different arbitrary 10-mer oligonucletide primers which showed polymorphism betwen tetraploid cultivars and haploid inducers used, three generated amplification products which seemed to be derived from the S. phureja parent. Our results indicate that chromosomes of dihaploids may not be pure S. tuberosum and the dihaploids may not be produced by parthenogenesis.

• 한국식품영양과학회지
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• 제29권4호
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• pp.606-614
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• 2000

Randomly Amplified Polymorphic DNA (RAPD) 기술을 이용하여 국내 식품으 로부터 분리한 Listeria sp. 분리균주에 대한 DNA polymorphism을 분석하고 유연환계를 비 교하며, 유용 marker를 개발할 목적으로 10가지 10-mer primer를 이용하여 PCR을 수행한 결과 5개의 primer(OPA-01, OP-26-01, OP-26-02, OPB-01, OP-26-10)가 선별되었고, 76개 의 DNA 단편이 증폭되었다. 이 중 OPA-01과 OP-26-10 promer에 의한 약 1.5 kb와 0.7 kb의 증폭 band는 모든 Listeria 분리균에서 관찰할수 있었으나, 이 증폭된 DNA 단편은 Listeria sp.에만 특이적인 것은 아니었다. NTSYS 프로그램을 이용해서 Listeria sp. 분리구 간의 유전적 유연관계를 알아본 결과 7개의 cluster로 나누어졌고 유사도는 대체로 0.54~ 0.93사이였으며, 특히, No.3과 No.20은 93%로 가장 높은 유사도를 나타내었고, No.7과 No.24 또는 No.7과 No.45는 54%로 가장 낮은 유사도를 나타내었다. 이러한 결과는 RAPS 기술을 이용하여 쉽게 Listeria sp.를 subspecies로 분류할 수 있음을 시사하였다.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
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• pp.66.1-66
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• 1996

• Journal of Ginseng Research
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• 제17권2호
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• pp.153-158
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• 1993

The study was carried out for comparison of variants and development of genetic markers using Randomly Amplified Polymorphic D사A (RAPD) analysis method. The ginseng variants used were as follows: Chungkyung-Chong, Hwangskoog-Chong, KG101 selected by the pureline selection method, and 6 kinds of Jakyung-Chong strains Uinjakyung, Jakyung-Chong 81783, Jakyung-Chong 847913, Jaky tong-Chong 79742, Jinjakyung of USSR, and Mimaki of Japan). Four of 10 RAPD primers showed the distinctive polymorphism among 9 ginseng variants and lines, and were selected for more detailed polymorphic analysis. The sequences of 4 selected primers were TGCCGAGCTG (Primer#2), AATCGGGCTG (#4), GAAACGGGTG (U7), and GTGACGTAGG (#8). All primers produced several common bands among the strains. However, when primer # 2 was applied, the electrophoregram showed the specific band at 1.8 kb region in Chungkyung-Chong, Hwangskoog- chong, and KG101, and 1 kb in the Jakyung-Chong 847913. In primer #4, 1.1 kb band was shown in Chungkyung-Chong, Hwangskoog-Chong, KG101, and Jakyung-Chong 79742. In primer # 7, 700 bp band was appeared in Jakyung-Chong 81783 and Jinjakyung of USSR In primer # 8, 800 bp band was observed only in Mimaki, comparing to another strains. When Similarity Index (SI) was calculated, Chungkyung-Chong and Hwngskoog-Chong, and Jakyung- chong 81783 and Jinjakyung of USSR showed the most close SI, 0.11 and 0.08, respectively. The data of KG101, which showed the SI of 0.13 with the group of Chungkyung-Chong and Hwangskoog-Chong, coincided with the fact that it was released from Hwangskoog-Chong by breeding process. The data of Jakyung strains indicated the significant variation among the strains. From these results, RAPD analysis method could be succesively applied to the classification and genetic analysis for breeding of Korean ginseng.

• Plant Biotechnology Reports
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• 제2권2호
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• pp.145-152
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• 2008

Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1535-1541
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• 2005

Five broiler chicken lines, namely HC, BPB2, CPB2, PB2 and UM1, involving in a selection program and differing in selection intensity and genetic background, were screened for randomly amplified polymorphic DNA (RAPD) polymorphism using 10 selected decamer primers. Nine primers amplified the genomic DNA, generating 200 to 2,500 bp and all detected polymorphism between lines. Out of 74 bands scored using these primers, 34 (50.0%) were found to be polymorphic. The number of polymorphic loci ranged from 3 to 6 with an average of 4.33. Lines differed considerably for within-population genetic similarity estimated by band frequency (WS = 93.55 to 99.25). Between-line genetic similarity estimates based on band sharing as well as on band frequency ranged from 71.35 to 86.45 and from 73.38 to 87.68, respectively. Lines HC and PB2 were the most closely related to the other, while BPB2 and CPB2 appeared to be more distant from each other. The between-line genetic distance based on both band sharing and band frequency revealed the similar trends as for Between-line genetic similarity. Based on BS and BF criteria, BPB2 and CPB2 as well as PB2 and UM1 lines can be merged to launch a new genetic group for further progress in biometrical objectives. A phylogenetic tree, derived using Nei's coefficient of similarity revealed the different pattern of genetic distance between lines.