• 한국육종학회지
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• 제42권4호
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• pp.365-373
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• 2010

TILLING (Targeting Induced Local Lesions IN Genomes) is broadly regarded as an excellent methodology for reverse genetics applications. Approximately 15,000 $M_3$ TILLING lines have been developed via the application of gamma-ray irradiation to rice seeds (cv. Donganbyeo), followed by subsequent selections. In an effort to evaluate the genetic diversity of the TILLING population, we have employed the AFLP multiple dominant marker technique. A total of 96 (0.64%) TILLING lines as well as Donganbyeo were selected randomly and their genetic diversity was assessed based on AFLP marker polymorphisms using 5 primer combinations. An average of 100.4 loci in a range of 97 to 106 was detected using these primer combinations, yielding a total of 158 (31.4%) polymorphic loci between Donganbyeo and each of the 96 lines. A broad range of similarity from 80% to 96% with an average of 89.4% between Donganbyeo and each of the 96 lines was also observed, reflecting the genetic diversity of the TILLING population. Approximately 28 polymorphic loci have been cloned and their sequences were BLAST-searched against rice whole genome sequences, resulting in 20 matches to each of the gene bodies including exon, intron, 1 kb upstream and 1 kb downstream regions. Six polymorphic loci evidenced changes in the coding regions of genes as compared to the rice pseudomolecules, 4 loci of which exhibited missense mutations and 2 loci of which exhibited silent mutations. Therefore, the results of our study show that the TILLING rice population should prove to be a useful genetic material pool for functional genomics as well as mutation breeding applications.

• 한국패류학회지
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• 제22권2호
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• pp.97-108
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• 2006

Genomic DNA samples isolated from geographical purple Washington clam (Saxidomus purpuratus) were obtained from two different regions in Korean Peninsula: Gunsan (Gunsan population; GSP), and Haeju (Haeju population; HJP), a collection area in the vicinity of the West Sea. The seven arbitrarily primers, OPA-07, OPA-09, OPA-18, OPA-20, OPC-03, OPC-06 and OPC-09 were shown to generate the total loci, loci observed per primer, shared loci by each population, specific, and polymorphic loci which could be clearly scored. We also generated the unique shared loci to each population and shared loci by the two populations in purple Washington clam. The size of the DNA fragments also varied wildly, from 50 to 2,400 bp. Here, 304 total loci were identified in the GSP purple Washington clam population, and 282 in the HJP: 91 polymorphic loci (29.9%) in the GSP and 47 (16.7) in the HJP. 198 shared loci, with an average of 28.3 per primer, were observed in the GSP population. The decamer primer OPA-07 generated the shared loci by the two populations, approximately 1,000 bp, between the two Saxidomus populations. The oligonucleotide primer OPC-03 also generated the shared loci by the two populations, approximately 500 bp and 1,000 bp, in GSP population from Gunsan and HJP population from Haeju. The other primer, OPC-06 generated the shared loci by two Gomphina populations (approximately 400 bp). The dendrogram, generated by seven reliable primers, indicates three genetic clusters. The dendrogram obtained by the seven primers indicates three genetic clusters: cluster 1 (GUNSAN 01-GUNSAN 02), cluster 2 (GUNSAN 03-GUNSAN 11), and cluster 3 (HAEJU 12-HAEJU 22). The genetic distance between the two geographical populations ranged from 0.043 to 0.506. Especially, the longest genetic distance displaying significant molecular differences, 0.506, was found to exist between individuals GUNSAN no. 11 of Gunsan and HAEJU no. 17 of Haeju.

• Asian-Australasian Journal of Animal Sciences
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• 제5권4호
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• pp.747-752
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• 1992

Protein polymorphism of native and red jungle fowls in Nepal was analyzed by electrophoresis. Blood samples were collected in the areas of Solu, Jomson road, Kathmandou, Pokhara and Low land. Out of 17 loci, polymorphism were found at nine loci in native fowls and at three loci in red jungle fowls. The proportion of polymorphic loci ($P_{poly}$) of native and red jungle fowls were $0.529{\pm}0.121$ and $0.176{\pm}0.095$, respectively. The five fowl populations in Nepal formed a different cluster from Sri Lankan and Bangladeshi fowl populations. When the gene frequencies of polymorphic loci were compared between the native fowl populations of Sri Lanka, Bangladesh and Nepal, $Amy-1^A$, $Es-1^A$ and $Akp-2^A$ genes showed inclination of south to north.

• Journal of Forest and Environmental Science
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• 제25권2호
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• pp.93-100
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• 2009

In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

• 한국산림과학회지
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• 제105권2호
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• pp.186-192
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• 2016

본 연구는 차세대 염기서열 분석방법을 이용하여 굴참나무의 microsatellite 마커를 개발하고 특성을 분석하기 위해 수행되었다. GS-FLX Titanium 차세대 염기서열 분석 장비를 이용하여 305,771개의 read를 얻었고, 117 Mbp의 데이터를 생산하였다. De novo assembly를 통하여 7,326개의 contig를 확보하였다. 크기가 500 bp 이상이 되는 contig는 2,921개로 나타났다. 그 중 microsatellite 영역을 포함하는 contig는 606개(20.75%)로 나타났으며, 총 microsatellite의 수는 911개로 확인되었다. 그 중 13개의 microsatellite 유전자좌에서 굴참나무 개체 간 다형성이 관찰되었다. 이들 microsatellite 유전자좌에 대하여 주왕산 집단에서 관찰된 유효 대립유전자수($A_e$)는 평균 4.966(2.439~7.515)로 나타났다. 평균 이형접합도 관측치($H_o$)와 평균 이형접합도 기대치($H_e$)는 각각 0.873(0.731~1.000)과 0.766(0.590~0.867)으로 나타났다. 다형성이 관찰된 모든 microsatellite 유전자좌에서 null 대립유전자는 관찰되지 않았으며, 마커 간 연관불평형은 나타나지 않았다. 따라서 본 연구에서 개발된 13개의 microsatellite 마커는 굴참나무 집단의 유전변이 분석에 유용할 것으로 사료된다.

• Journal of Plant Biology
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• 제39권3호
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• pp.223-229
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• 1996

The genetic variation in Korean evening primorse (Oeothera odorata L.) populations was examiend to estimate the level of allozyme variation within populatons using starch gel electrophoresis. 7 of 13 loci (Adh, Est-1, Est-2, Mdh-2, Pgd-2, Pgm-1, and Idh) revealed (Ps=43.2%) were polymorphic. The mean number of alleles per locus (A) and polymorphic locus (Ap) for populations were 1.64 and 2.46, respectively. The effective number of alleles (Aep) within populations relatively was low ranging from 1.08 to 1.22 with a mean of 1.14. Within populations, the mean number of allele per polymorphic loci (Ap) was 2.46, the mean number of alleles per locus (A) was 1.64, and the mean genetic diversity was 0.093. About 2.7% of the total allozyme diversity resided among populations (Mean GST=0.0274). FIS, a measure of the deviation from random mating within 13 populations, was relative low (mean FIS=0.03636). The indirect estimate of gene flow, based on the mean GST, was high (Nm=8.88). Estimates of gene flow were consistent with low levels of genetic differentiation among populations.

• International Journal of Industrial Entomology and Biomaterials
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• 제35권2호
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• pp.97-99
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• 2017

Eleven polymorphic microsatellite loci were developed and characterized for Dorcus hopei in this study. The number of alleles varied from 2 to 21. The observed heterozygosity and expected heterozygosity ranged from 0.1058 to 0.9744 and 0.0997 to 0.8941, respectively. Two loci showed low polymorphism, while the rest were highly polymorphic. Six loci deviated from Hardy-Weinberg Equilibrium. The set of markers will provide effective tools for examining the population genetic structures and be helpful for managing wild population in D. hopei.

• International Journal of Industrial Entomology and Biomaterials
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• 제28권2호
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• pp.39-50
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• 2014

In order to understand the diversity and genetic relationships of silkworm strains preserved in Korea, we genotyped 78 Bombyx mori strains (Bombycidae: Lepidoptera) originating from Japan, using eight polymorphic microsatellite loci. We obtained per-locus allele numbers ranging from 5 to 16 (with an average value of 9.1), per-locus observed heterozygosity ranging from 0.13 to 1.00, and per-locus polymorphic information content ranging from 0.36 to 0.77, indicating that some loci are highly variable. Phylogenetic analysis with the eight concatenated microsatellite loci showed no clustering based on known strain characteristics and origin. Nineteen strain-specific apomorphic alleles, which discriminated 16 of the 78 silkworm strains, were obtained from eight loci. These strain-specific alleles can thus be utilized for routine discrimination of strains from Japan, without any further typing of other loci. Homozygotes were also observed at some loci (27 of 118 genotypes), which can also be used to discriminate several strains by typing a few loci. These results showed that eight microsatellite loci described herein were sufficiently variable to discriminate among the 78 silkworm strains we examined, and may be useful for future investigations of this economically important species.

• 한국발생생물학회지:발생과생식
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• 제10권4호
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• pp.227-238
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• 2006

한반도의 동쪽에 위치해 있는 삼척(venus clam from Samcheok; VCS)과 원산(venus clam from Wonsan; VCW) 지역에서 채취된 민들조개(Gomphina aequilatera)에서 genomic DNAs(gDNAs)를 분리 추출하였다. 증폭산물은 primer agarose 전기영동법에 의해서 생성되었고, EtBr에 의해서 염색된 이후에 자외선에 의해서 확인되었다. 150 bp에서 2,400 bp에 해당되는 shared loci, polymorphic 및 specific loci를 얻기 위해서 BION-21, BION-23, BION-25, BION-27, BION-29, BION-31 및 BION-33와 같은 7개의 primer를 사용하였다. 본 연구에서 7개의 primer는 VCS 민들조개 집단에서 147개의 polymorphic loci(147/954 loci, 15.41%)와 VCW 집단에서 274개의 polymorphic loci(274/996 loci, 27.51%)를 확인하였다. 이것은 VCS 민들조개 집단에서 보다 VCW 집단에서 더 높은 유전적 변이를 나타내고 있다는 것을 제시하고 있다. 특히 BION-21 primer에 의해서 나타난 700 bp는 민들조개 2개 집단에서 공통적으로 확인되었으며, 이러한 것은 집단이나 종을 확인할 수 있는 marker로서 활용이 가능할 것이다. 이러한 특이한 primer는 개체, 종 및 집단에서 서로 다른 DNA 다형성을 나타내며, 개체나 집단을 확인하는 데 유용하다는 것을 알 수 있다. 2개 민들조개 집단의 개체들을 비교해 보았을 때 SAMCHEOK no. 03와 WONSAN no. 22에서 가장 긴 유전적 거리(0.696)를 나타내었다. 3개의 genetic groupings and dendrogram을 포함한 complete linkage cluster analysis을 통해서 볼 때 지리적 거리가 있었지만 삼척과 원산 2 민들조개 집단의 개체 정체성과 다소 가까운 친척관계를 확인시켜 주었다. 분자적인 표지인자로부터 얻어진 종내 분류와 clustering analyses은 패각 크기, 패각 형태 및 패각 색깔과 같은 형태적인 형질을 기초한 재래적인 종 분류를 지원하고 있다. 따라서 위에서 언급된 바와 같이 RAPD 분석은 VCS 민들조개 집단이 VCW 집단과 어느 정도 차이가 있다는 것을 확인시켜 주었다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권4호
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• pp.453-457
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• 2005

The genetic variations in three major river populations viz. the Halda, the Jamuna and the Padma of the Indian major carp, Catla catla were analyzed by Random Amplified Polymorphic DNA (RAPD) markers. Four decamer primers were used for amplifying DNA of 10 individuals from each population. The proportion of polymorphic loci and the gene diversity estimates were 59.4 and 0.20 for the Halda, 37.5 and 0.14 for the Jamuna and 46.9 and 0.16 for the Padma populations respectively indicating the existence of a relatively high level of genetic variation in the Halda river population. The inter-population similarity indices, gene flow and genetic distance values indicated that the Jamuna-Padma population pair of catla was genetically closer than the Halda-Jamuna and the Halda-Padma population pairs in compliance with the geographical distances among them. The coefficient of gene differentiation ($G_{ST}$=0.13) reflects some degree of genetic differentiation among three populations of catla studied. The data suggest that the RAPD technique could be used to discriminate different river populations of catla.