鲤鱼微卫星分子标记的筛选

微卫星 (ｍｉｃｒｏｓａｔｅｌｌｉｔｅ)是近十几年来发展起来的一种新的分子标记 ,它是指以少数几个核苷酸 ( 1～ 6个 )为单位多次重复的简单序列 ,以双核苷酸重复最为常见 ,而其中又以 (ＣＡ/ＧＴ) ｎ 居多。由于微卫星在真核生物基因组中是随机分布的 ,而作为分子遗传标记又有着非常高的多态性和共显性 ,因此在构建遗传连锁图谱时备受青睐 (Ｂｒｏｏｋｅｔａｌ ,1994 )。目前 ,在人类和多种动物中已经构建了以微卫星为主的遗传连锁图谱。但在鲤鱼等水产动物的遗传连锁图谱中 ,微卫星分子标记还较少 (孙效文和梁利群 ,2 0 0 0 )。为了摸索…

Isolation of Microsatellite Markers in the Common Carp (Cyprinus carpio)

A partial common carp genomic library was constructed.45 positive clones were isolated from screening about 2000 clones of the genomic library with a (CA)15 probe labelled at the 5　' end with γ32P-ATP.Sequencing of these clones was performed with automated sequencer，and 22 microsatellites were isolated.17 primers were designed based on unique sequences flanking each motif with the software Primer3.PCR on Cyprinus carpio haematopterus was carried out with these primers，and all gave expected bands.Annealing temperature of these primers was between 50℃ and 53℃.