中国水产科学2011，Vol.18Issue(1)：194-201,8.DOI:10.3724/SP.J.1118.2011.00194
三疣梭子蟹微卫星富集文库的构建与群体遗传分析
Construction on enriched microsatellite library and characterization of microsatellite markers from swimming crab
摘要
Abstract
Thirty microsatellite markers of swimming crab Portunus trituberculatus were isolated and characterized by screening SSR-enriched library. Genomic DNA were digested by HaeⅢ, then DNA fragments containing microsatellites were captured by a piece of nylon membrane(Hybond N+) bounded with the probe combinations of (AC)15 and (AG)15. After transformation to Escherichia coli strain DH5α, the clones were regularly re-arrayed in a new agar plate with the density of about 300 clones per plate and were screened with (AC)15 and (AG)15 probes labeled by DIG luminescent Detection Kit system(Roche). One hundred and fifty clones were randomly selected for sequencing. One hundred and five primer pairs were designed using the software Primer Premier 5.0, of which 56 pairs could amplify distinct PCR products. Polymorphism of these distinct loci was assessed using 30 swimming crab individuals, and the results showed that the 30 loci were all polymorphic. The values of Ho, He and PIC varied from 0.222 2 to 1.000 0, from 0.436 7 to 0.909 9 and from 0.350 to 0.892, respectively. These microsatellite markers would be useful for the studies of genome mapping, QTL, parentage determination and population genetics for this species. [Journal of Fishery Sciences of China, 2011, 18(1): 194-201]关键词
三疣梭子蟹/富集文库/菌落原位杂交/微卫星标记/遗传多样性分类
农业科技引用本文复制引用
李晓萍,刘萍,宋协法..三疣梭子蟹微卫星富集文库的构建与群体遗传分析[J].中国水产科学,2011,18(1):194-201,8.基金项目
国家863高技术研究发展计划项目(2006AA10A406) （2006AA10A406）
国家自然科学基金项目(30871933,40806067) （30871933,40806067）
中央级公益性科研院所基本科研业务费专项资金项目(2008-ts-11). （2008-ts-11）
