ISSR-PCR技术在对虾中的应用初步研究

Preliminary study on the application of inter-simple sequence repeats (ISSR)-PCR technique in Chinese shrimp ( Fenneropenaeus chinensis)

采用 ISSR(Inter Simple Sequence Repeats)技术对中国对虾 (Penaeus chinensis)进行了 PCR扩增。优化了 PCR反应体系和反应参数 ,对 PCR产物进行聚丙烯酰胺凝胶电泳和银染检测。从 10 0条 ISSR引物中筛选了 4 0条有清晰产物的引物 ,每条引物检测到的位点数从 1到 19不等 ,平均每条引物可检测到位点数为 7.7个。实验发现 :中国对虾简单重复序列区主要由两碱基循环组成。通过分析 ISSR- PCR技术本身的原理 ,探讨了该技术相对于同工酶检测和 RAPD技术在遗传多样性分析中的优势所在 。

Inter Simple Sequence Repeats (ISSR) was the first time to be applied to detect the genetic variation in Fenneropenaeus chinensis. The reaction system for ISSR was optimized; denatured polyacrylamide gel electrophoresis (PAGE) and silver staining were employed in isolating the amplified fragments. 40 ISSR primers screened from 100 primers could generate clear bands. Each primer yielded from 1 to 19 bands with the average of 7 7 bands. The results demonstrated that the simple sequence tandem repeats (STRs) were mainly composed of dinucleotede repeats. Based on its unique characters, ISSR technique can detect more genetic loci than isozyme and has higher stability than RAPD which are two main techniques applied to assess the genetic diversity at present. ISSR has potential not only on the analyzing the genetic markers but also on the genetic maps construction of Fenneropenaeus chinensis.