一步3′RACE快速构建鸡MnSOD全长cDNA克隆

Rapid Construction of Full-length MnSOD cDNA of Chickens by One-step 3′RACE

将触减PCR与3′cDNA末端快速扩增(rapidamplificationofcDNAends,RACE)技术进行结合,仅用一条特异性引物和一条通用引物,成功地实现了从3′末端cDNA库对鸡含锰超氧化物歧化酶(manganese-containingsu peroxidedismutase,MnSOD)全长cDNA的一步3′RACE快速构建。与常规使用的末端PCR或亚克隆方法相比,该法具有快速、省时、经济和特异性好的优点。

RACE(rapid amplification of cDNA ends) is a popular technique to rapidly obtain the full-length cDNA. After obtaining the 3′cDNA and 5'cDNA fragments with a overlapped region by 3′RACE and 5′RACE, the full-length cDNA could be generated by end-to-end PCR or subcloning. In this study, 3′RACE combined with touch-down PCR was successfully used for the rapid construction of full-length MnSOD cDNA of chickens. Compared with the conventional end-to-end PCR or subcloning, this method, called one-step 3′RACE, is fast, economical and highly specific. It especially fits the rapid construction of full-length cDNA by RACE method.