摘要: |
采用 CAP3 软件对 NCBI 上的 5296 条缢蛏 ESTs 序列进行了微卫星特征分析。 结果表明, 经拼接、去冗得到非冗余 EST 序列 3453 条, 含 SSR 位点的 EST 序列 267 条, 共 307 个 SSR 位点, 检出率为 8.89%, 平均每 6.83kb 出现 1 个 SSR 位点。设计了 40 对 EST-SSR 引物并进行 PCR 扩增, 29对引物能扩增出理想的 PCR 产物, 其中多态性引物 14 对。利用 14 对多态性引物分析了乐清湾缢蛏遗传多样性, 共检测到等位基因数(Na)61 个, 每个位点的等位基因数为 2—12 个。二核苷酸、三核苷酸和四核苷酸重复是最主要的重复类型, 分别占 15.96%、37.13%和 35.50%。乐清湾缢蛏群体观测杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)分别为 0.569、0.490 和 0.449, 表明乐清湾缢蛏遗传多样性较丰富。 |
关键词: 缢蛏, 表达序列标签(EST), 简单重复序列(SSR) |
DOI:10.11693/hyhz201201022022 |
分类号: |
基金项目:温州市科技兴海项目, S20080019 号;浙江省近岸水域生物资源开发与保护重点实验室开放基金, J2010009 号;浙江省海水养殖重点科技创新团队资助, 2010R50025 号;国家贝类产业技术体系浙江综合试验站资助, CARS-48 号 |
附件 |
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CHARACTERIZATION, DEVELOPMENT AND UTILIZATION OF EST-DERIVED MICROSATELLITES IN SINONOVACULA CONSTRICTA |
LIU Bo1,2, SHAO Yan-Qing, TENG Shuang-Shuang, CHAI Xue-Liang, XIAO Guo-Qiang
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1.Zhejiang Mariculture Research Institute;2.Zhejiang Key Laboratory of Exploitation and Reservation of Coastal Bio-resource
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Abstract: |
A set of 5296 expressed sequence tags (ESTs) of Sinonovacula constricta from NCBI were screened using CAP3.0 software. After assembly, a total of 3453 non-redundant EST sequences were mined and yielded 267 non-redundant sequences which contained 307 putative SSRs accounted for 8.89% of the total number of EST sequences. This is equivalent to one EST-SSR per 6.83kb of sequence. Di-, Tri- and Tetranucleotide repeats EST-SSRs were dominant, counting for 15.96%, 37.13% and 35.50% respectively. Forty primers were designed for partial EST-SSRs and used for PCR amplification with 29 primer pairs showing amplifications while 14 were polymorphic. These were used to conduct SSR analyses of genetic variations of the 40 individual samples of S. constricta collected in Yueqing Bay. A total of 61 alleles were detected, with the number of alleles ranging from 2 to 12. The observed heterozygosity (Ho), expected heterozygosity (He) and polymorphism information content (PIC) were 0.569, 0.490 and 0.449, respectively. All of these parameters indicate that the S. constricta of Yueqing Bay rich genetic variation. |
Key words: Sinonovacula constricta, Expressed sequence tags (EST), Simple sequence repeats (SSR) |