太湖新银鱼微卫星位点的分离与序列特征分析

为了开发太湖新银鱼（Neosalanx taihuensis）的微卫星分子标记，采用生物素标记探针（AC）12、（AAC）10、（AAG）10和（GATA）8对其微卫星位点进行了筛选，并对其序列特征进行了分析。共获得490个微卫星序列，筛选的总效率为78．09％。筛选出725个微卫星位点，其中完美型592个，占81．66％；混合型82个，占11．31％；非完美型51个，占7．03％。探针（AC）12富集到的微卫星重复次数大多集中在14～26次，最高44次；探针（AAC）10和探针（AAG）10富集得到的微卫星重复次数主要集中在8～20次，最高42次；探针（GATA）8富集得到的微卫星重复次数一般在6～15次，最高32次。探针（AC）12、（AAC）10、（AAG）10和（GATA）8的杂交效率分别为85．19％、60．19％、5．16％和10．64％。筛选得到的725个微卫星位点中，二碱基重复位点390个（53．79％），三碱基重复位点284个（39．17％），四碱基重复位点47个（6．48％），五碱基重复位点 1个（0．14％）和六碱基重复位点3 个（0.42％）。根据二碱基重复核心序列进行引物设计，对抚仙湖24尾太湖新银鱼样本进行 PCR 扩增，电泳结果显示部分微卫星位点有较高的多态性，同时本研究获得的三碱基、四碱基、五碱基和六碱基重复位点也可为长重复单元微卫星引物的开发提供基础。

Isolation and Sequence Characterization of Microsatellite loci in Neosalanx taihuensis

Microsatellite markers from Neosalanx taihuensis were screened using (AC)12, (AAC)10, (AAG)10 and (GATA)8 oligonucleotide probes respectively. A total of 490 microsatellite sequences were identified and total efficiency of screening was 78.09%. There were 592 (81.66%) perfect motifs, 82 (11.31%) compound motifs, and 51(7.03%) imperfect motifs among these sequences. After enrichment using the probe (AC) 12 the microsatellites were found to be repeated on average 14 to 26 times, the largest of which was repeated was 44 times. Separating microsatellite using the probes (AAC)10 and (AAG)10 resulted in repeat numbers of 8~20 and a maximum of 42 repetitions; the microsatellite repeat number from the probe (GATA)8 averaged around 6 to 15 times, with a maximum of 32 repetitions. We screened 390 two base repeat loci, 284 three base repeat loci, 47 four base repeat loci, 1 five base repeat locus and 3 six base repeat loci in this study. Screening these microsatellite molecular loci could lead to important further research on N.taihuensis.