程序化设计的简并引物克隆半滑舌鳎CYP17基因  

Programming Design of Degenerate Primers and Cloning Half-Smooth Tongue Sole Cynoglossus semilaevis CYP17 Gene

作　　者：陈彩芳[1] 温海深[1] 何峰[1] 董双林[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2009年第6期1213-1218,共6页Periodical of Ocean University of China

基　　金：国家"十一五"科技支撑计划课题(2006BAD09A01)资助

摘　　要：采用1种新的简并引物设计方法——CODEHOP法来克隆半滑舌鳎CYP17基因。该法结合NCBI、Blastp、BlockMaker、CODEHOP等生物信息学资源和DNAMAN、Oligo6.0等生物软件,程序性设计半滑舌鳎CYP17基因的CODEHOP引物,从半滑舌鳎卵巢组织中成功克隆了CYP17基因,该基因片段与其它物种的CYP17家族具有较高的同源性,经鉴定,该基因属于整个脊椎动物的CYP17基因群,尤其是鱼类亚群。实验结果表明,程序性设计的简并引物较传统简并引物特异性更高,假阳性率少,更有利于实验成功。new method of designing degenerate primers-CODEHOP is introduced to clone half-smooth tongue sole Cynoglossus semilaevis CYP17 gene. With Bioinformatics Resources such as NCBI, Blastp, Block- Maker.CODEHOP and biological software such as DNAMAN,Oligo6.0, the CODEHOP primer is programming designed, and finally we succeeded to clone the gene from the ovaries of half-smooth tongue sole Cynoglossus semilaevis. The fragment of the gene has high identities with the CYP17 family of other species, and it is clustered into the CYP17 gene group of all the other vertebrates, especially into the fish subgroup. The experiment results showed that compared with traditional degenerate primers, programming designed primers had more Specific, lower positive rate, and more benefit for accomplishing the experiment.

关 键 词：半滑舌鳎 简并引物 CYP17 

分 类 号：S917.4[农业科学—水产科学] Q7[生物学—分子生物学]