• Biotechnology and Bioprocess Engineering:BBE
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• 제5권1호
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• pp.23-26
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• 2000

In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of $85.3\%$. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed $83.0\%$ of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short micro satellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, MC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found.

• 생명과학회지
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• 제16권3호
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• pp.382-386
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• 2006

고양이의 혈통 등록을 위한 개체식별 및 친자판정을 목적으로 microsatellite DNA다형을 조사한 결과 다음과 같은 성적을 얻었다. 고양이 20두를 대상으로 microsatellite DNA다형의 대립유전자를 조사한 본 연구에서는 관찰된 대립유전자의 수는 $3{\sim}8$개 (평균 5.5개)이며 marker별 대립유전자는 FCA005 142bp (0.3750), FCA026 148 bp (0.5500), FCA075 136 bp (0.5000), FCA105 191 bp (0.4250), FCA224 156 bp (0.7750), FCA229 166 bp (0.6500), FCA240 163 bp (0.3000), FCA293 185 bp (0.5000), FCA453 186 bp (0.5500), FCA651 134 bp (0.6750) 대립유전자가 높은 빈도로 관찰되었다. Expected heterozygosity와 PIC는 각각 $0.390{\sim}0.827$(평균 0.639), $0.357{\sim}0.780$(평균 0.581)으로 나타났고 FCA240의 marker는 PIC value가 0.70 이상이었다. 또한 PE는 $0.076{\sim}0.444$으로서 10개 marker를 조합시 total PE는 0.9441로 관찰되었다. 10개의 microsatellite DNA다형 좌위를 가지고 친자관계를 분석한 결과 8두 중에서 1두(12.50%)가 모순으로 판정되었다. 또한 국내에서 사육중인 고양이의 개체식별 및 친자판정에 microsatellite marker를 활용할 수 있을 것으로 사료된다.

• 한국양식학회지
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• 제19권3호
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• pp.205-209
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• 2006

현재 우리나라에서 종묘방류량이 많은 넙치의 유전적 다양성을 파악하기 위하여 양식산 어미 암컷 31마리, 수컷 52마리로 총 83마리로부터 생산된 종묘의 유효어미수와 근교계수를 microsatellite DNA marker 7개를 이용하여 추정하였다. 어미집단과 종묘집단의 대립유전자수는 어미집단에 비하여 하루 동안 채란한 E1 종묘집단에서 23.5%가 감소하였고, 이틀 동안 채란한 E2 종묘집단에서는 17.6%가 감소하였다. 유전자 동일성검사 결과, 산란에 관여한 어미수는 E1 종묘집단에서 총 23마리였으며 이중 암컷 9마리, 수컷 14마리였고, E2 종묘집단에서는 35마리로 암컷 15마리, 수컷 20마리였다. 유효 어미집단크기(Ne)를 추정하기 위해 산란에 관여한 실제 어미의 암수의 비율을 보정하면, Ne는 E1 종묘집단에서 21.9마리, E2 종묘집단에서 34.3마리로 추정되었다. 이에 따라 근교계수는 El 종묘집단이 0.023, E2종묘집단은 0.015로서 E2종묘집단에서 낮게 나타났다. 이와 같은 결과는 수산생물의 다양성을 보존하기 위한 FAO의 권고 기준보다 높게 나타남에 따라 유전적 다양성을 향상시키는 방류용 종묘생산 방안이 필요할 것으로 나타났다.

• 생명과학회지
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• 제19권2호
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• pp.174-178
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• 2009

본 연구는 2006년 8월경에 어획된 서해, 남해 및 동해 멸치집단을 분석하기 위하여 6종류의 microsatellite로 하였다. 서해멸치의 경우, 샘플수 72마리에 대한 allele 범위가 19-41로 평균 28.5를 보였다. 특히 EJ9 locus에서 평균보다 약 1.4배 많은 41를 나타내었다. 남해멸치의 평균 allele는 24.5로 서해보다는 적었고, EJ2, EJ9, EJ27.1 loci는 평균보다 높은 29-37의 범위를 보였다. 동해멸치는 평균 allele가 25.0으로 EJ35를 제외하면 대부분의 loci에서 평균 이상을 보였다. 그러나 서해, 남해 및 동해멸치의 allele 빈도율은 대부분 0.24 이하로 나타났다. 또한 Hobs보다 Hexp에서 0.5 정도 높은 값을 보였으나 유의성은 없었다(p>0.05). 유전적 다양성도 0.9 이상으로 매우 높은 값을 보였다. 6종류의 microsatellite에 대한 지역간의 유전적 차이 및 거리는 0.258과 0.019로 유의적으로 차이는 없었다 (p>0.05). 따라서 서해, 남해 및 동해 멸치계군은 유전적으로 동일한 집단을 형성하고 있는 것으로 판단된다.

• 생명과학회지
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• 제33권8호
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• pp.623-631
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• 2023

본 연구는 차세대 염기서열 분석법(NGS)을 사용하여 벤자리의 microsatellite 마커를 개발하고, 개발한 마커를 사용하여 벤자리 집단의 유전학적 특성을 분석하기 위해 수행되었다. 차세대 염기서열 분석 장비인 Illumina Hiseq X ten을 사용하여 총 402,244,934개의 read들을 얻어 assembly를 실행한 결과, 전체의 약 0.33%에 해당하는 1,320,995개의 read들이 assembly 되었으며, 이 read들의 총 길이는 705,613,658 bp로 확인되었다. 크기가 640 bp 이상이 되는 contig는 952,326개로 나타났으며, 이 중 microsatellite 영역을 포함하는 contig를 151개(0.016%)로 1차 선별하고, PCR 증폭 여부를 통해 microsatellite 후보 34개를 2차로 선별하였다. 그 중 벤자리 집단의 마커로서 유용한 15개의 microsatellite 마커를 최종 선택하였다. 새로 개발한 15개의 microsatellite 마커를 사용하여 벤자리 집단을 대상으로 분석한 결과, 관찰된 유효 대립유전자 수(N_A)는 평균 12(6~25)로 나타났다. 평균 관측치 이형접합도(H_O)와 평균 기대치 이형접합도(H_E)는 각각 0.750(0.530-0.873)와 0.793 (0.647-0.895)으로 나타냈으며, 이는 해산어의 평균 값인 0.79와 유사한 수치를 나타내었다. 따라서 본 연구에서 개발한 15개의 microsatellite 마커는 벤자리 집단의 유전학적 특성 분석에 유용할 것으로 사료된다.

• Fisheries and Aquatic Sciences
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• 제15권2호
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• pp.151-155
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• 2012

We used nine microsatellite DNA markers to estimate genetic variation among wild and cultured populations of the sea squirt Halocynthia roretzi. The loci were polymorphic, with 6-32 alleles, and allelic richness ranged from 6.0 to 26.1 in each population. The wild and the cultured populations had similar mean heterozygosities ($H_O$ and $H_E$), allele numbers, and allelic richness. One cultured population with softness syndrome had a lower mean in the observed heterozygosity ($H_O$ = 0.57) and higher mean inbreeding coefficient ($F_{IS}$ = 0.261) than any other populations. This suggests that the loss of genetic variation in the diseased population might be due to increased inbreeding. A neighbor-joining tree and pairwise population estimates of $F_{ST}$ showed moderate genetic differentiation between the wild and the cultured populations. Additionally, the softness syndrome population was genetically divergent from wild populations, but it was genetically close to the cultured populations.

• Asian-Australasian Journal of Animal Sciences
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• 제20권12호
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• pp.1783-1790
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• 2007

The genetic diversity of domestic quail and two wild quail species, Japanese （Coturnix coturnix）and Common quail (Coturnix japonica), found in China was studied using microsatellite DNA markers. According to a comparison of the corresponding genetic indices in the three quail populations, such as Polymorphism Information Content (PIC), Mean Heterozygosity ($\bar{H}$) and Fixation Index, wild Common quail possessed rich genetic diversity with 4.67 alleles per site. Its values for PIC and $\bar{H}$ were the highest, 0.5732 and 0.6621, respectively. Domestic quail had the lowest values, 0.5467 and 0.5933, respectively. Wild Japanese quail had little difference in genetic diversity from domestic quail. In addition, from analyses of the fuzzy cluster based on standard genetic distance, the similarity relationship matrix coefficient between wild Japanese quail and domestic quail was 0.937, and that between wild Common quail and domestic quail was 0.783. All of these results showed that the wild Japanese quail were closer to the domestic quail for phylogenetic relationship than wild Common quail. These results at the molecular level provide useful data about quail's genetic background and further supported the hypothesis that the domestic quail originated from the wild Japanese quail.

• Animal Bioscience
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• 제35권9호
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• pp.1314-1326
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• 2022

Objective: Alongside the rise of animal-protection awareness in Taiwan, the public has been paying more attention to dog genetic deficiencies due to inbreeding in the pet market. The goal of this study was to isolate novel microsatellite markers for monitoring the genetic structure of domestic dog populations in Taiwan. Methods: A total of 113 DNA samples from three dog breeds-beagles (BEs), bichons (BIs), and schnauzers (SCs)-were used in subsequent polymorphic tests applying the 14 novel microsatellite markers that were isolated in this study. Results: The results showed that the high level of genetic diversity observed in these novel microsatellite markers provided strong discriminatory power. The estimated probability of identity (P_(ID)) and the probability of identity among sibs (P_(ID)sib) for the 14 novel microsatellite markers were 1.7×10^-12 and 1.6×10^-5, respectively. Furthermore, the power of exclusion for the 14 novel microsatellite markers was 99.98%. The neighbor-joining trees constructed among the three breeds indicated that the 14 sets of novel microsatellite markers were sufficient to correctly cluster the BEs, BIs, and SCs. The principal coordinate analysis plot showed that the dogs could be accurately separated by these 14 loci based on different breeds; moreover, the Beagles from different sources were also distinguished. The first, the second, and the third principal coordinates could be used to explain 44.15%, 26.35%, and 19.97% of the genetic variation. Conclusion: The results of this study could enable powerful monitoring of the genetic structure of domestic dog populations in Taiwan.

• 한국수산과학회지
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• 제50권6호
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• pp.806-811
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• 2017

A multiplex PCR system comprising 14 microsatellite markers was developed for genotyping analysis of the sea cucumber Stichopus japonicus. A total of 286 samples were used to evaluate genetic polymorphisms and forensic parameters of the microsatellite loci. In a single PCR reaction, all 14 loci were uniformly amplified and a total of 269 alleles were identified. The AJ19024 locus had the largest number of alleles (46), and its discriminatory power and exclusion power were 0.99 and 0.76, respectively. The fewest alleles (8) were present at the Psj2575 locus, which provided the lowest discriminatory power (0.81) and exclusion power (0.20). The mean number of alleles, mean heterozygosity, mean discrimination power and mean exclusion power per locus were 19.21, 0.70, 0.93, and 0.46, respectively. The combined matching probability for the 14 loci was $9.64{\times}10^{-19}$, and the combined power of exclusion was 0.999995. Thus, the forensic parameters evaluated in the present study demonstrated the utility of our multiplex PCR system for biological tracing methods, such as individual identification and paternity testing, in the sea cucumber.

• International Journal of Industrial Entomology and Biomaterials
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• 제45권2호
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• pp.49-55
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• 2022

A total of 20 mulberry varieties preserved in Korea were typed for eight polymorphic microsatellite loci. We obtained 6 to 15 alleles per locus with an average value of 10.6, per-locus observed heterozygosity ranging from 0.35 to 1.00, and per-locus polymorphic information content (PIC) ranging from 0.61 to 0.87, indicating that most loci are highly variable. Phylogenetic analysis using the eight microsatellite loci was sufficiently suitable for classifying 20 mulberry varieties preserved in Korea. A total of 160 variety-specific apomorphic alleles were obtained from eight loci discriminated 20 mulberry varieties. These variety-specific alleles from this analysis are expected to be useful for the discrimination of other mulberry varieties. Furthermore, a substantial number of homozygote loci, represented by 60 among 180 alleles in eight loci were found. These results collectively suggest that these microsatellite locus primers are potentially crucial molecular markers for the eventual classification of mulberry varieties that are preserved as hundreds in Korea.