兰州鲇G-SSR和EST-SSR在鲇形目和鲤形目鱼类中的通用性研究

Transferability Analysis of Silurus lanzhouensis G-SSR and EST-SSR Markers in Siluriformes and Cypriniformes Fishes

为研究兰州鲇G-SSR和EST-SSR在鲇形目和鲤形目鱼类中的通用性,以12种不同鱼类为试验材料,分别采用改进的酚-氯仿法结合DNA产物纯化试剂盒法、传统酚-氯仿法、试剂盒法提取其尾鳍基因组DNA,对提取的DNA进行琼脂糖凝胶电泳、紫外分光光度计检测。结果表明,改进的酚-氯仿法结合DNA产物纯化试剂盒法提取的鱼类基因组DNA在纯度和稳定性方面明显优于传统酚-氯仿法、试剂盒法,且电泳条带清晰、整齐、明亮,操作简单,耗时短,便于快速、批量提取。利用兰州鲇14对G-SSR引物和21对EST-SSR引物在12种鱼类中进行跨目通用性分析,结果显示,兰州鲇G-SSR和EST-SSR在鲇形目、鲤形目鱼类的通用率分别为63.10%、32.14%和61.11%、44.44%,表明随着物种间亲缘关系变远,其通用性随之降低。

In order to investigate the transferability of Silurus lanzhouensis G-SSR and EST-SSR in Siluriformes and Cypriniformes,12 different fishes were used as research objectives to extract genomic DNA by using improved phenol-chloroform method,which combined with DNA purification kit,traditional phenol-chloroform method and DNA extraction kit,respectively.The genomic DNA was detected by agarose gel electrophoresis and ultraviolet spectrophotometer. The results showed that the genomic DNA extracted by the improved phenol-chloroform method was obviously superior to the other two methods in purity and stability,and the electrophoresis bands were very clear,neat and bright. The method had the advantages of simple manipulating,short time-consuming,speediness and batch extraction. Furthermore,14 G-SSRs and21 EST-SSRs of Silurus lanzhouensis were selected to analyze the transferability of 12 Siluriformes and Cypriniformes fishes. The results indicated that the transferable rates of 14 G-SSRs and 21 EST-SSRs in Siluriformes and Cypriniformes fishes were 63. 10%,32. 14% and 61. 11%,44. 44% respectively,revealing that the transferability decreased with the increase of genetic relationship among species.