• 제목/요약/키워드: polymorphic microsatellite

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Molecular Characterization of Hallikar Breed of Cattle Using Microsatellite Markers

  • Kumar, S. Naveen;Jayashankar, M.R.;Nagaraja, C.S.;Govindaiah, M.G.;Saravanan, R.;Karthickeyan, S.M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권5호
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    • pp.622-626
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    • 2006
  • Molecular characterization of Hallikar, the native cattle breed of Karnataka, was undertaken using 19 cattle specific, highly polymorphic microsatellite markers recommended by FAO. The genomic DNA was subjected to PCR amplification and alleles were resolved through six per cent denaturing PAGE with a 10 bp DNA ladder followed by silver staining. Genotyping of animals was done based on allele size. The number of alleles ranged from three to nine with allele sizes ranging from 102 bp to 294 bp. These alleles were distributed in the frequency range between 0.0306 and 0.8673 in the population. The mean observed number of alleles was $6.368{\pm}1.4225$. The mean observed and expected heterozygosities were $0.7515{\pm}0.1734$ and $0.7850{\pm}0.1381$, respectively. The high heterozygosity observed implies presence of higher genetic variability within Hallikar breed. The PIC (Polymorphism Information Content) values ranged from 0.2322 (ETH152) to 0.8654 (ETH225). The percentage of polymorphic loci obtained was 100 as all the 19 microsatellite markers were found to be polymorphic. Except for ETH152, all the other loci had high PIC values, indicating that these markers are highly informative for characterization of Hallikar breed. The population was tested for Hardy-Weinberg equilibrium at 19 microsatellite loci, and at 74 per cent of the loci the population was found to be in disequilibrium.

New polymorphic microsatellite markers for the endangered fern Ceratopteris thalictroides (Parkeriaceae)

  • CHO, Won-Bum;HAN, Eun-Kyeong;KWAK, Myounghai;LEE, Jung-Hyun
    • 식물분류학회지
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    • 제48권2호
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    • pp.129-133
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    • 2018
  • Ceratopteris thalictroides is a semi-aquatic fern with a circumtropical distribution. Because this species is designated internationally on the IUCN Red List as requiring at least some concern, Korean populations are of great concern for the species' long-term survival, as they are at the northern limit of the species distribution. To establish an effective conservation strategy for those populations at the genetic level, we used the Mi-Seq platform to develop three sets of 25 polymorphic microsatellite markers for C. thalictroides, which is endangered in Korea. In populations sampled from Busan and Gochang, the number of alleles ranged from 2 to 13 (average of 5.64), and plants presented an expected heterozygosity of 0.000 to 0.860. These markers will be useful for evaluating the genetic status and conserving Korean populations of C. thalictroides more effectively.

Development and characterization of eleven microsatellite markers for a popular pet stag beetle, Dorcus hopei (Coleoptera, Lucanidae) using paired-end Illumina shotgun sequencing

  • Han, Taeman;Kim, Seung-Hyun;Park, In Gyun;Park, Haechul
    • 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.

Development of Novel Microsatellite Markers for Strain-Specific Identification of Chlorella vulgaris

  • Jo, Beom-Ho;Lee, Chang Soo;Song, Hae-Ryong;Lee, Hyung-Gwan;Oh, Hee-Mock
    • Journal of Microbiology and Biotechnology
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    • 제24권9호
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    • pp.1189-1195
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    • 2014
  • A strain-specific identification method is required to secure Chlorella strains with useful genetic traits, such as a fast growth rate or high lipid productivity, for application in biofuels, functional foods, and pharmaceuticals. Microsatellite markers based on simple sequence repeats can be a useful tool for this purpose. Therefore, this study developed five novel microsatellite markers (mChl-001, mChl-002, mChl-005, mChl-011, and mChl-012) using specific loci along the chloroplast genome of Chlorella vulgaris. The microsatellite markers were characterized based on their allelic diversities among nine strains of C. vulgaris with the same 18S rRNA sequence similarity. Each microsatellite marker exhibited 2~5 polymorphic allele types, and their combinations allowed discrimination between seven of the C. vulgaris strains. The two remaining strains were distinguished using one specific interspace region between the mChl-001 and mChl-005 loci, which was composed of about 27 single nucleotide polymorphisms, 13~15 specific sequence sites, and (T)n repeat sites. Thus, the polymorphic combination of the five microsatellite markers and one specific locus facilitated a clear distinction of C. vulgaris at the strain level, suggesting that the proposed microsatellite marker system can be useful for the accurate identification and classification of C. vulgaris.

Genetic diversity analysis of fourteen geese breeds based on microsatellite genotyping technique

  • Moniem, Hebatallah Abdel;Zong, Yang Yao;Abdallah, Alwasella;Chen, Guo-hong
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권11호
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    • pp.1664-1672
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    • 2019
  • Objective: This study aimed to measure genetic diversity and to determine the relationships among fourteen goose breeds. Methods: Microsatellite markers were isolated from the genomic DNA of geese based on previous literature. The DNA segments, including short tandem repeats, were tested for their diversity among fourteen populations of geese. The diversity was tested on both breeds and loci level and by mean of unweighted pair group method with arithmetic mean and structure program, phylogenetic tree and population structure were tested. Results: A total of 108 distinct alleles (1%) were observed across the fourteen breeds, with 36 out of the 108 alleles (33.2%) being unique to only one breed. Genetic parameters were measured per the 14 breeds and the 9 loci. Medium to high heterozygosity was reported with high effective numbers of alleles (Ne). Polymorphic information contents (PIC) of the screened loci was found to be highly polymorphic for eleven breeds; while 3 breeds were reported moderately polymorphic. Breeding coefficient ($F_{IS}$) ranged from -0.033 to 0.358, and the pair wise genetic differentiation ($F_{ST}$) ranged from 0.01 to 0.36 across the fourteen breeds; for the 9 loci observed and expected heterozygosity, and Ne were same as the breeds parameters, PIC of the screened loci reported 6 loci highly polymorphic and 3 loci to be medium polymorphic, and $F_{IS}$ ranged from -0.113 to 0.368. In addition, genetic distance estimate revealed a close genetic distance between Canada goose and Hortobagy goose breeds by 0.04, and the highest distance was between Taihu goose and Graylag goose (anser anser) breed by 0.54. Conclusion: Cluster analyses were made, and they revealed that goose breeds had hybridized frequently, resulting in a loss of genetic distinctiveness for some breeds.

Establishment of an Individual Identification System Based on Microsatellite Polymorphisms in Korean Cattle (Hanwoo)

  • Yoon, Du-Hak;Kong, Hong-Sik;Oh, Jae-Don;Lee, Jun-Heon;Cho, Byung-Wook;Kim, Jong-Dae;Jeon, Ki-Jun;Jo, Chang-Yun;Jeon, Gwang-Joo;Lee, Hak-Kyo
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권6호
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    • pp.762-766
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    • 2005
  • This study was conducted to establish an individual identification system comprising of 19 microsatellite markers located on different bovine autosomes. The markers were typed on 257 animals from five cattle breeds. In total, 112 alleles were detected from the genotyping of 19 microsatellite markers. The average heterozygosities ranged from 0.292 to 0.824 and the polymorphic information content (PIC) ranged from 0.274 to 0.817 in Hanwoo. We found that there were differences in allele frequencies in Hanwoo when compared with other cattle breeds. The calculated cumulative power of discrimination (CPD) was 99.999% when nine microsatellite loci were used for analysis in the individual identification system. Also the matching probability, the probability that two unrelated animals would show the same genotypes, was estimated to be $0.44{\times}10^{-9}$. Therefore, the nine markers used in this study will be used for individual identification in two million Hanwoo individuals.

New polymorphic microsatellite markers in the Korean mi-iuy croaker, $Miichthys$ $miiuy$, and their application to the genetic characterization of wild and farmed populations

  • An, Hye-Suck;Kim, Eun-Mi;Lee, Jang-Wook;Kim, Dae-Jung;Kim, Yi-Cheong
    • Animal cells and systems
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    • 제16권1호
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    • pp.41-49
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    • 2012
  • Eighteen new polymorphic microsatellite markers were developed for the Korean mi-iuy croaker ($Miichthys$ $miiuy$, Perciformes, Sciaenidae), and allelic variability was compared between a wild population in Mokpo, Korea, and a hatchery population in Tongyeong, Korea. All loci were amplified readily and demonstrated allelic variability, with the number of alleles ranging from 5 to 37 in the wild population, and from 4 to 12 in the farmed population. The average observed and expected heterozygosities were estimated, respectively, to be 0.74 and 0.78 in the hatchery population samples, and 0.79 and 0.86 in the wild samples. These results indicate lower genetic variability in the hatchery population compared with the wild population, and significant genetic differentiation between the wild population and the hatchery samples ($F_{ST}$=0.058, P<0.001). These microsatellite loci may be valuable for future population genetic studies, monitoring changes in the genetic variation within stocks in a commercial breeding program, conservation genetics, and molecular assisted selective breeding of the mi-iuy croaker in the future.

Development of Microsatellite Markers to Distinguish South Korean and Chinese Ginseng

  • Ahn, Chang-Ho;Kim, Boo-Bae;Yoon, Eui-Soo;Choi, Yong-Eui
    • 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.

Development and Characterization of 10 Polymorphic Microsatellite Loci in the Korean Endemic Freshwater Fish Iksookimia koreensis, and Their Cross-species Amplification in the Endemic I. longicorpa

  • Kwan, Ye-Seul;Kim, Hyo-Jin;Lee, Bit-Na;Won, Yong-Jin
    • Animal Systematics, Evolution and Diversity
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    • 제33권2호
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    • pp.136-139
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    • 2017
  • The genus Iksookimia (Actinopterygii: Cypriniformes: Cobitidae) is a bottom-dwelling freshwater loaches, which are well-known as their endemism and high geographic variation. However, population genetic relationships among Iksookimia spp. have remained unclear due to a shortage of genetic markers that can be applied generally in the genus. Here, we developed high-resolving microsatellite markers using I. koreensis and I. longicorpa as representatives of Iksookimia species because of their wide distribution range and phylogenetic position. Ten of polymorphic microsatellite loci were isolated from Iksookimia koreensis and were successfully cross-amplified in I. longicorpa. The mean number of observed alleles per locus was about 10.4 (range, 2-17) for I. koreensis and about 13.2 (range, 2-24) for I. longicorpa. The loci, IK03 and IK08, deviated from the Hardy-Weinberg equilibrium in I. koreensis, after applying the Bonferroni correction. The microsatellite markers obtained in the present study will be useful to evaluate population genetic structure and to establish conservation strategies for I. koreensis and related Iksookimia species.

차세대 염기서열 분석법을 이용한 방어(Seriola quinqueradiata)의 microsatellite 마커의 개발 및 유전적 특성 분석 (Development and Genetic Diversity Analysis of Microsatellite Markers Using Next-generation Sequencing in Seriola quinqueradiata)

  • 동춘매;이미난;김은미;박중연;김군도;노재구
    • 생명과학회지
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    • 제30권3호
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    • pp.291-297
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    • 2020
  • 본 연구는 차세대 염기서열 분석법(NGS)을 이용하여 방어의 microsatellite 마커를 개발하고, 개발된 마커를 이용하여 방어 집단의 유전적 특성을 분석하기 위해 수행되었다. 차세대 염기서열 분석 장비인 Illumina Hiseq2500를 이용하여 총 28,873,374개의 read들을 얻어 assembly를 수행한 결과, 전체의 약 1.6%에 해당하는 466,359개의 read들이 assembly 되었으며, 이 read들의 총 길이는 7,247,216,874 bp로 확인되었다. 크기가 518 bp 이상이 되는 contig는 30.729개로 나타났으며, 이 중 microsatellite 영역을 포함하는 contig 132개(0.43%)를 1차로 선별하고, PCR 증폭 여부 및 유전자형 분석을 통해 microsatellite 후보 60개를 2차로 선별하였다. 그 중 방어집단의 마커로서 유용한 15개의 microsatellite 마커를 선택하였다. 방어집단을 대상으로 개발된 15개의 microsatellite 마커로 분석한 결과, 관찰된 유효 대립유전자수(NA)는 평균 18.5(11~30)로 나타났다. 평균 관측치 이형접합도(HO)와 평균기대치 이형접합도(HE)는 각각 0.812(0.431~0.972)와 0.896(0.782~0.949)으로 나타났다. 다형성이 관찰된 모든 microsatellite 마커 간의 연관불평형은 나타나지 않았으며, 해산어의 평균 HE 값인 0.79 이상의 수치를 나타내었다. 따라서 본 연구에서 개발된 15개의 microsatellite 마커는 방어 집단의 유전적 다양성 분석에 유용할 것으로 사료된다.