| 磁珠富集法制备翘嘴红鲌微卫星分子标记 |
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| 引用本文： | 张义凤,孙效文,鲁翠云,梁利群,徐伟,李勇,刘晓锋.磁珠富集法制备翘嘴红鲌微卫星分子标记[J].农业生物技术学报,2008,16(4). |
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| 作者姓名： | 张义凤 孙效文 鲁翠云 梁利群 徐伟 李勇 刘晓锋 |
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| 作者单位： | 1. 中国水产科学研究院黑龙江水产研究所,哈尔滨,150070;大连水产学院生命科学与技术学院,大连,116023
2. 中国水产科学研究院黑龙江水产研究所,哈尔滨,150070 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)，国家公益性计划 |
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| 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的方式开发翘嘴红鲌基因组微卫星资源。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。
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| 关 键 词： | 翘嘴红鲌 微卫星 磁珠富集 |
| 收稿时间： | 2007-11-11 |
| 修稿时间： | 2008-1-21 |
Microsatellite Enrichment by Magnetic Beads in Erythroculter ilishaeformis(Bleeker) |
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| ZHANG Yi-feng,SUN Xiao-wen,LU Cui-yun,LIANG Li-qun,XU Wei,LI Yong,LIU Xiao-feng.Microsatellite Enrichment by Magnetic Beads in Erythroculter ilishaeformis(Bleeker)[J].Journal of Agricultural Biotechnology,2008,16(4). |
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| Authors: | ZHANG Yi-feng SUN Xiao-wen LU Cui-yun LIANG Li-qun XU Wei LI Yong LIU Xiao-feng |
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| Abstract: | According to the hadro-affinity 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 1400 colonies. Sequencing of 288 positive colonies confirmed that it contained 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. 243 pairs of primers were designed with the software Primer 3.0 .In addition, 50 pairs of primers were composed and screened. 30 pairs were used successfully to amplify special fragemnts, among which 12 pairs were polymorphism within species |
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