不同对虾种间共用微卫星DNA引物的研究

采用已发表的斑节对虾的30对微卫星引物,对部分微卫星引物在中国对虾、凡纳对虾、日本对虾中的通用性进行了研究。通过所建立的降温PCR(Touchdown PCR)方法对3种对虾的DNA进行PCR扩增,筛选出3对引物在3种对虾中均能产生清晰谱带。根据上述结果适当调整PCR反应体系中各试剂的量及反应条件,达到了理想的结果。利用这3对引物对3种对虾进行遗传标记分析,结果显示,PCR扩增产物条带清晰,特异性强,每对引物在3种对虾间扩增出的特异性片段大小几乎一致。研究表明,3种对虾基因组间存在一定程度的相似性,引物W15-16、W21-22和W45-46可直接应用于中国对虾、凡纳对虾、日本对虾、斑节对虾分子标记的基因组比较作图,也为图谱克隆法提供了新的思路。

Common Microsatellite Primers for Molecular Markers in Different Shrimp Species

Using 30 pairs of microsatellite primers from Penaeus monodon , common microsatellite primers were studied in marine shrimp Fenneropenaeus chinensis , Litopenaeus vannamei and Marsupenaeu japonicus. DNAs extracted from the 3 shrimp species were amplified by "Touchdown" PCR. 3 pairs of common microsatellite primers were screened and produced clear fragments. Optimal results were obtained by optimizing quantity of reagents and reaction conditions. Under the optimal conditions, PCR was performed for analyzing the genetic markers of the 3 species. As a result , clearly distinctive fragments were produced and were almost the same size by the same microsatellite primers for the 3 species, which indicated that the 3 shrimp species have certain extents of comparability, and the 3 pairs of common primers W15-16,W21-22 and W45-46 can be applied for the molecular markers on genome comparative mapping and map-based gene cloning of the 3 shrimp species.