磁珠富集法制备翘嘴红鲌微卫星分子标记  

Microsatellite Enrichment by Magnetic Beads in Erythroculter ilishaeformis(Bleeker)

作　　者：张义凤[1] 孙效文[1] 鲁翠云[1] 梁利群[1] 徐伟[1] 李勇[1] 刘晓锋[1] 

机构地区：[1]中国水产科学研究院黑龙江水产研究所

出　　处：《农业生物技术学报》2008年第4期610-615,共6页Journal of Agricultural Biotechnology

基　　金：国家重点基础研究发展规划(973)(No．2004CB117405);国家公益性计划资助项目(No．2005DIB4J024)资助。

摘　　要：根据生物素与链霉亲和素的强亲和性原理,采用生物素.磁珠吸附微卫星富集法与传统的放射性同位素杂交法相结合开发翘嘴红鲌(Erythroculter ilishaeformis)基因组微卫星资源。结果表明,从1400个菌落中筛选出613个阳性克隆,占38.31%。将288个进行测序,得到262个(94.44%)含有微卫星序列的克隆,其中含有微卫星座位280个。完美型204个,占72.86%;非完美型49个,占17.5%;复合型27个,占9.64%。设计获得243对微卫星引物,合成50对经PCR筛选,结果30对引物可扩增出特异性条带。12对具有多态性。According to the hadro-affmity principle of biotin and streptavidin, microsatellite DNAs were isolated from Erythroculter ilishaeformis genome by combining biotin capture method and radioactive labeling hybridization. In this experiment, 613 （38.31%） positive colonies were obtained through 1 400 colonies. Sequencing of 288 positive colonies confirmed that 262 of which contained microsatellite sequence clone and 280 microsatellite loci. From these sequences, 204 repeat motifs （about 72.86%） were perfect, 49 repeat motifs （about 17.5%） were imperfect, 27 repeat motifs （about 9.64%） were compound. Two hundred and forty-three pairs of primers were designed with the software Primer 3.0, 50 pairs of primers were composed and screened. Results showed that 30 pairs were used successfully to amplify special fragments, and 12 pairs of them were polymorphism within species.

关 键 词：翘嘴红鲌 微卫星 磁珠富集 

分 类 号：S188[农业科学—农业基础科学]