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细角螺微卫星DNA富集文库构建及特征分析 被引量:1
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作者 李清荟 陈晓姣 +3 位作者 黎中宝 曹媛钰 陈丽娜 戴刚 《海洋科学》 CAS CSCD 北大核心 2013年第4期1-5,共5页
采用生物素磁珠富集法,用生物素标记的(GT)15和(CT)15两种探针与细角螺基因组MseI酶切的300~1200bp片段杂交,杂交复合物与链霉亲和素磁珠结合,捕获含有重复序列的微卫星片段。最后将磁珠捕获到的重复序列与PMD19-T载体连接后克隆到DH5... 采用生物素磁珠富集法,用生物素标记的(GT)15和(CT)15两种探针与细角螺基因组MseI酶切的300~1200bp片段杂交,杂交复合物与链霉亲和素磁珠结合,捕获含有重复序列的微卫星片段。最后将磁珠捕获到的重复序列与PMD19-T载体连接后克隆到DH5α中构建微卫星基因组文库。通过PCR检测出354个阳性克隆,从中随机选取248个片段大于500bp的阳性克隆进行测序,结果显示,在220个成功测序的阳性克隆中共获得278个微卫星序列,其中完美型171个,占61.5%;非完美型81个,占29.1%;复合型26个,占9.4%。除探针使用的GT和CT的重复序列外,还筛选到三碱基GTT、TGG、GAA、CAA;四碱基TCTA、ACAG、TAGA、GTGA、GTCT、GATA及五碱基TTTTG的重复序列。在278条序列中共有82条可以设计引物。 展开更多
关键词 细角螺(Hemifusus ternatanus) 磁珠富集 微卫星 构建 DNA文库
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Analysis of the genetic diversity and differentiation of Fenneropenaeus penicillatus populations using AFLP technology 被引量:1
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作者 张桂玲 曹媛钰 +4 位作者 黎中宝 陈锦 赵斌丽 雷光高 王展林 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2012年第3期440-445,共6页
Fenneropenaeus penicillatus (redtail shrimp) is an important marine commercial animal in China. Recently, its resources have been depleted rapidly as a result of, for example, over-exploitation and environmental deg... Fenneropenaeus penicillatus (redtail shrimp) is an important marine commercial animal in China. Recently, its resources have been depleted rapidly as a result of, for example, over-exploitation and environmental degradation of spawning grounds. Therefore, we analyzed the genetic diversity and differentiation of nine wild populations of F. penicillatus of China (Ningde, Lianjiang, Putian, Xiamen, Quanzhou, Zhangpu, Dongshan, Nanao, and Shenzhen populations) by amplified fragment .!ength polymorphism (AFLP) technology, to provide genetic information necessary for resource protection, rejuvenation, artificial breeding, and sustainable use of the resource. Eight AFLP primer pairs were used for amplification, and 508 bands were detected among the populations. The results show that the percentage of polymorphic loci (P) ranged from 41.34% to 63.58%; the Nei's gene diversity (H) of the populations was 0.119 4-0.230 5; and Shannon's Information Index (/) was 0.184 1-0.342 5. These genetic data indicate that the genetic diversity of F. penicillatus was high. The genetic differentiation coefficient (GsT=0.216 2) and gene flow (Nm=1.812 4) show that there was a high level of genetic differentiation and a moderate level of gene flow among populations. More studies on the genetic differentiation mechanism of F. peniillatus along the south-eastem coast of China need to be conducted to find more effective scientific protection strategies for the conservation ofF. penicillatus genetic resources. 展开更多
关键词 Fenneropenaeus penicillatus AFLP genetic diversity genetic differentiation
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