用CODEHOP法设计简并引物克隆草鱼多聚免疫球蛋白受体基因片段

摘要

The gene of polymeric immunoglobulin receptor(pIgR)of grass carp Ctenopharyngodon idellus was cloned by a pair of degenerate primers designed by CODEHOP, and by combined bioinformatics including NCBI, Blast and biological software such as Primer Premier 5.0 and DNAMAN, and the homology and phylogenetic tree of the clone of pIgR gene were analyzed in the PCR products.The results showed that degenerate primers designed by CODEHOP were characterized by specificity and benefit for the experiment accomplishing.pIgR gene of grass carp was found to clustered into pIgR gene group in all the other vertebrates, especially into the fish subgroup, with the maximal homology(84%)with common carp Cyprinus carpio.%为克隆草鱼Ctenopharyngodon idellus多聚免疫球蛋白受体pIgR基因, 本研究中借助于NCBI、 Blast等数据库及Primer Premier 5.0、 DNAMAN等生物软件, 利用CODEHOP法设计了简并引物, 并对克隆基因进行同源性比较和系统发生分析.结果表明: 用CODEHOP法设计的简并引物特异性强, 试验成功率高;草鱼pIgR基因属于整个脊椎动物的pIgR基因群, 与鲤Cyprinus carpio同源性最高, 为84%.