• 식물분류학회지
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• 제41권4호
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• pp.299-314
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• 2011

분자 마커의 선택은 집단유전학의 연구방법을 결정하는 중요한 고려사항으로, 현재까지 동식물의 집단유전학 연구에는 알로자임, RAPD, RFLP, AFLP, microsatellite, SNP, ISSR 등이 개발되어 주로 사용되고 있다. 이 중 microsatellite는 핵뿐만 아니라 엽록체, 미토콘드리아와 같은 세포소기관의 게놈상에 매우 풍부하게 존재하며, 핵에서 유래된 microsatellite는 높은 다형성을 보이는 공우성 마커로 집단 구조 및 유전적 다양성 연구에서 최근 선호된다. Microsatellite는 보통 1~6 bp의 짧은 서열이 반복된 것으로 각각의 유전자좌에 특화된 프라이머를 사용하여 증폭한다. Microsatellite는 PCR 반응으로 쉽게 유전자형을 분석할 수 있는 장점이 있으나, 종 특이적으로 개발되고 계통적으로 매우 가까운 근연종에게만 적용될 수 있는 단점이 있다. 따라서, 야생식물의 경우 microsatellite 개발에 필요한 게놈 정보가 부족하고 신규 개발비용이 많이 소요되어 적용이 쉽지 않았으나, 점차 개발비용이 낮아지고 있어, 야생식물을 대상으로 한 microsatellite 연구들이 증가하고 있는 추세이다. 따라서, 본 논문에서는 야생식물의 microsatellite를 이용한 분석 기초를 마련하고자 microsatellite 마커의 다양한 개발 및 분석 방법, 진화 모델 및 적용 분야에 대해 소개하고, 유전자형 결정시 잘못된 결론을 도출할 가능성이 높은 부분에 대한 사항들을 지적하여 야생식물의 microsatellite를 이용한 집단유전학적 분석에 도움을 주고자 하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권10호
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• pp.1329-1333
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• 2004

To isolate the microsatellites from the chromosomal DNA of the Korean cattle (Hanwoo) and to use those for the genetic selection, four bacteriophage genomic libraries containing the chromosomal DNA of six Hanwoo steers showing the differences in meat quality and quantity were used. Screening of the genomic libraries using $^{32}P-radiolabeled 5'-({CA})_{12}-3$nucleotide as a probe, resulted in isolation of about 3,000 positive candidate bacteriophage clones that contain $(CA)_n$-type dinucleotide microsatellites. After confirming the presence of microsatellite in each positive candidate clone by Southern blot analysis, the DNA fragments that include microsatellite and flanking sequences possessing less than 2 kb in size, were subcloned into plasmid vector. Results from the analysis of microsatellite length polymorphism, using twenty-two PCR primers designed from flanking region of each microsatellite DNA, demonstrated that 208 and 210 alleles of HW-YU-MS#3 were closely related to the economic traits such as marbling score, daily gain, backfat thickness and M. longissimus dorsi area in Hanwoo. Interestingly, HW-YU-MS#3 microsatellite was localized in bovine chromosome 17 on which QTLs related to regulation of the body fat content and muscle ypertrophy locus are previously known to exist. Taken together, the results from the present study suggest the possible use of the two alleles as a DNA marker related to economic trait to select the Hanwoo in the future.

• Asian-Australasian Journal of Animal Sciences
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• 제16권11호
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• pp.1555-1559
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• 2003

The objective of the present study was to explore associations between five microsatellites linked to $Fec^B$ and $FecX^I$ genes and litter size in Small Tail Han sheep. The polymorphisms of five microsatellite loci, OarAE101, BM1329, BMS2508, TGLA54 and TGLA68 were detected in 244 ewes of Small Tail Han sheep. Analysis of association between three microsatellite loci (BMS2508, BM1329 and OarAE101) located in the 10 cM region covering the $Fec^B$ gene (Booroola gene) and litter size in Small Tail Han sheep indicated that BMS2508 had significant effect on litter size in the second parity (p<0.05), but no significant effect on litter size in the first parity (p>0.05), while the other two microsatellite loci had no significant effect on litter size in both the first and the second parity in Small Tail Han sheep (p>0.05). At microsatellite locus BMS2508, least squares means in the second parity of genotypes 101/111 and 99/109 were significantly higher than those of genotypes 99/99, 99/101, 99/111 and 99/115 (p<0.05); least squares mean in the second parity of genotype 101/111 was significantly higher than that of genotypes 109/111 and 111/111 (p<0.05). Results of this study also indicated that two microsatellite loci (TGLA54 and TGLA68) that confined the 28.7 cM region covering the $FecX^I$ gene (Inverdale gene) did not affect litter size in both the first and the second parity in Small Tail Han sheep significantly (p>0.05). The information found in the present study is very important for improving the reproductive performance in sheep breeds by marker assisted selection.

• Mycobiology
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• 제45권4호
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• pp.401-408
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• 2017

Colletotrichum gloeosporioides is an economically important fungal pathogen causing substantial yield losses indifferent host plants. To understand the genetic diversity and molecular epidemiology of this fungus, we have developed a novel, high-resolution multi-locus microsatellite typing (MLMT) method. Bioinformatic analysis of C. gloeosporioides unannotated genome sequence yielded eight potential microsatellite loci, of which five, CG1 $(GT)_n$, CG2 $(GT1)_n$, CG3 $(TC)_n$, CG4 $(CT)_n$, and CG5 $(CT1)_n$ were selected for further study based on their universal amplification potential, reproducibility, and repeat number polymorphism. The selected microsatellites were used to analyze 31 strains of C. gloeosporioides isolated from 20 different host plants from India. All microsatellite loci were found to be polymorphic, and the approximate fragment sizes of microsatellite loci CG1, CG2, CG3, CG4, and CG5 were in ranges of 213-241, 197-227, 231-265, 209-275, and 132-188, respectively. Among the 31 isolates, 55 different genotypes were identified. The Simpson's index of diversity (D) values for the individual locus ranged from 0.79 to 0.92, with the D value of all combined five microsatellite loci being 0.99. Microsatellite data analysis revealed that isolates from Ocimum sanctum, Capsicum annuum (chili pepper), and Mangifera indica (mango) formed distinct clusters, therefore exhibited some level of correlation between certain genotypes and host. The developed MLMT method would be a powerful tool for studying the genetic diversity and any possible genotype-host correlation in C. gloeosporioides.

• 한국축산식품학회지
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• 제27권3호
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• pp.357-362
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• 2007

Nine brand populations of Hanwoo cattle were characterized using 11 microsatellite DNA markers. The studied populations were: Ansung, Yangpyang, DaeGwanryeng, Palkongsangkangwoo, Hoengseong, Jangsu, Sumjinkang, Hadong, Nam-hae. The observed heterozygosity, expected heterozygosity, and polymorphism information content were calculated. Allele frequencies were calculated and used for the characterization of each brand population and to study their genetic relationships. Genetic distances were estimated using Nei's DA genetic distance and the resultant DA matrix was used in the construction of phylogenetic trees. The NJ tree showed that Ansung and Yangpyang, Sumjinkang and Jangsu, Namhae and Ha-Dong are closely related and are considered to have undergone genetic exchange within the same locale. This study will contribute to the local Hanwoo brand industry.

• 한국산림과학회지
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• 제98권5호
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• pp.568-575
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• 2009

Korean wild and forest cultivated ginseng has long been accepted as high medicinal values compared to field cultivated ginseng. Owing to the high price of Korean wild ginseng, Chinese wild and forest cultivated ginseng were smuggled and sold as Korean wild and forest cultivated ginseng. Therefore, an efficient method is required to distinguish Korean ginseng from Chinese ginseng. Microsatellites, simple sequence repeats (SSRs), are highly polymorphic loci present in DNA that consist of repeating units of base pairs. Thus SSR markers are highly advantageous for detection of small genetic variances of intra-species. In the present study, we constructed a microsatellite-enriched genomic library from South Korean wild Panax ginseng. After sequence analysis of 992 randomly picked positive colonies, 126 (12.7%) of the colonies were found to contain microsatellite sequences, and 38 primer pairs were designed. By polymorphism assessment using 36 primer pairs, 4 primers (PG409, PG450, PG491, and PG582) were shown to be polymorphic to distinguish the South Korean ginseng from the Chinese ginseng. These 4 microsatellite markers will provide powerful tools to authenticate South Korean ginseng from Chinese ginseng.

• Asian-Australasian Journal of Animal Sciences
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• 제31권2호
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• pp.167-179
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• 2018

Objective: The aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations. Methods: Novel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms. Results: 14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses. Conclusion: Based on this study's analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.

• 원예과학기술지
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• 제31권5호
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• pp.580-589
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• 2013

국내에서 최근에 유통되고 있는 고추 170품종을 대상으로 microsatellite 마커를 이용하여 DNA 프로파일 데이터베이스를 구축하기 위하여 품종식별력이 높은 분자표지의 선정 및 이를 활용한 품종 식별력 및 유전적 유사도 검정 등에 대한 연구를 수행하였다. 고추 형태적 특성이 다른 11품종을 302개의 microsatellite 마커로 검정하여 24개의 다형성이 높은 마커를 선정한 다음 170품종에 대한 DNA 프로파일 데이터베이스를 구축하였다. 고추 170품종을 24개의 microsatellite 마커로 분석하였을 때 마커당 평균 대립유전자수는 6.83개로 나타났고, 최소 2개부터 15개까지 다양한 분포를 나타내었다. PIC 값의 경우 0.324-0.873 범위에 속하였으며 평균값은 0.673으로 높게 나타났다. Microsatellite 마커의 대립유전자를 이용하여 고추 170품종에 대한 계통도를 작성하였을 때 과실의 형태에 따라 3개의 그룹으로 크게 나누어졌으며 모든 품종이 microsatellite 마커의 유전자형에 의해 식별이 되는 것으로 나타났다. 이 연구결과에 의해 개발된 고추 품종별 DNA 프로파일 데이터베이스는 품종보호출원 품종의 선 DNA 검정을 통한 대조품종 선정, 구별성, 균일성, 안정성의 재확인에 매우 유용하게 이용되어질 수 있어 향후, 품종보호권 강화 등에 크게 기여할 수 있을 것으로 사료된다.

• Journal of Animal Science and Technology
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• 제54권6호
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• pp.401-409
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• 2012

The objective of this study was to determine genetic variation of five Korean native chicken lines using 30 microsatellite (MS) markers, which were previously recommended by ISAG (International Society for Animal Genetics). The initial study indicated that two microsatellite markers, MCW0284 and LEI0192, were not amplified in these lines and excluded for further analysis. Twenty eight microsatellite markers were investigated in 83 birds from five Korean native chicken lines. The identified mean number of alleles was 4.57. Also, the expected, observed heterozygosity (He, Ho) and polymorphism information content (PIC) values were estimated in these markers and they ranged from 0.31~0.868, 0.145~0.699, and 0.268~0.847, respectively. The results were used for the discrimination of five chicken lines using genetic distance values and also neighbor-joining phylogenetic tree was constructed. Based on the He and PIC values, eighteen markers are enough for the discrimination of these Korean native chicken lines for the expected probability of identity values among genotypes of random individuals (PI), random half sibs ($PI_{half-sibs}$) and random sibs ($PI_{sibs}$). Taken together, these results will help the decision of conservation strategies and establishment of traceability system in this native chicken breed. Also, the use of ISAG-recommended microsatellite markers may indicate that the global comparison with other chicken breeds is possible.

• Asian-Australasian Journal of Animal Sciences
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• 제29권2호
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• pp.176-183
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• 2016

A study was conducted to assess the genetic diversity among Simmental Cross cattle in West Sumatra using microsatellite DNA markers. A total of 176 individual cattle blood samples was used for obtaining DNA samples. Twelve primers of microsatellite loci as recommended by FAO were used to identify the genetic diversity of the Simmental Cross cattle population. Multiplex DNA fragment analysis method was used for allele identification. All the microsatellite loci in this study were highly polymorphic and all of the identified alleles were able to classify the cattle population into several groups based on their genetic distance. The heterozygosity values of microsatellite loci in this study ranged from 0.556 to 0.782. The polymorphism information content (PIC) value of the 12 observed loci is high (PIC>0.5). The highest PIC value in the Simmental cattle population was 0.893 (locus TGLA53), while the lowest value was 0.529 (locus BM1818). Based on the genetic distance value, the subpopulation of the Simmental Cross-Agam and the Simmental Cross-Limapuluh Kota was exceptionally close to the Simmental Purebred thus indicating that a grading-up process has taken place with the Simmental Purebred. In view of the advantages possessed by the Simmental Cross cattle and the evaluation of the genetic diversity results, a number of subpopulations in this study can be considered as the initial (base) population for the Simmental Cross cattle breeding programs in West Sumatra, Indonesia.