拟穴青蟹(Scylla paramamosain)微卫星位点开发-基于(AG)12探针杂交  

NEW MICROSATELLITE LOCI IN THE MUD CRAB SCYLLA PARAMAMOSAIN DEVELOPED BY HYBRIDIZATION TO (AG)12 BIOTIN-LABELED PROBE

作　　者：宋忠魁[1,2] 聂振平[1] 谢达[1] 王芳宇[3] 

机构地区：[1]广西海洋研究所广西海洋生物技术重点实验室北海536000 [2]钦州学院化学与生物系钦州535000 [3]衡阳师范学院生命科学系衡阳421008

出　　处：《海洋与湖沼》2013年第1期171-176,共6页Oceanologia Et Limnologia Sinica

基　　金：广西科学基金项目,0991275号;广西科学研究与技术开发计划项目,11107012-11号;广西海洋生物技术重点实验室主任基金课题,0801号

摘　　要：采用FIASCO(Fast Isolation by AFLP sequences containing repeats)技术和磁珠富集方法开发拟穴青蟹微卫星位点。结果表明,400bp以下长度的序列含微卫星的概率超过400bp以上长度的序列;从二碱基重复类型到五碱基重复类型,完美型微卫星的概率在增加。利用设计的28对引物扩增一个供试群体,其中12对引物能稳定扩增且片段大小基本符合理论长度,10个微卫星位点表现出高度多态性,9个微卫星位点显著偏离Hardy-Weinberg平衡(P<0.05),两组两两位点间存在连锁不平衡现象(P<0.0042,经Bonferroni法校正)。Micro-checker分析揭示其中的5个微卫星位点可能存在无效等位基因。排除混合微卫星位点的引物对以及扩增位点PIc值在0.5以下的引物对,8对引物能用于拟穴青蟹群体遗传学等研究。FIASCO （Fast Isolation by AFLP Sequences Containing repeats） technique and magnetic bead enrichment method were used to develop microsatellite loci of the mud crab Scylla paramamosain. The results showed that more microsatellite sequences distributed within the range of 400bp in length and the probabilities of perfect microsatellite increased from di-to pentamer repeated types. 28 primer pairs were designed to amplify a sampling population, while 12 polymorphic loci were amplified, whose length conformed to theory size basically. Ten loci displayed polymorphic highly and nine loci were deviated from Hardy-Weinberg equilibrium （HWE） significantly （P〈0.05）. Significant linkage disequi- librium was detected in two pairwise loci （P〈0.0042, corrected by Bonferroni method）. Micro-Checker analysis suggested that five loci were involved in PCR amplification of null alleles. Regardless of the primer pairs of microsatellite mixture （containing two microsatellite sites or more）, plus some loci with PIC value lower than 0.5, eight primer pairs proved use-ful for the related population genetic analysis.

关 键 词：FIASCO技术 磁珠富集法 拟穴青蟹 微卫星 

分 类 号：Q953.5[生物学—动物学] S917.4[农业科学—水产科学]