悦目金蛛丝腺SMART RACE cDNA文库的构建与鉴定

Construction and Characterization of SMART RACE cDNA Library of Silk Gland in Argiope amoena

运用SMART技术构建了悦目金蛛丝腺SMARTRACEcDNA文库。经检测,文库所含全长cDNA的长度主要集中在500bp～2000bp之间;把双链cDNA通过T/A克隆、随机挑选阳性克隆并测序后,得到1条752bp的全长cDNA序列。以该文库为模板,用依据这条全长cDNA序列设计的基因特异性引物与接头引物进行RACE,3’RACE和5’RACE的产物拼接后的全长序列与上述全长cDNA序列一致。结果表明,该文库适于用RACE方法从中分离在悦目金蛛丝腺中表达基因的全长cDNA。本文还对SMART技术的特点和局限性进行了讨论。

A SMART RACE eDNA library of silk gland in Argiope amoena was constructed successfully with SMART technique. The result showed that the length of the full-length eDNAs in the library was pooled mainly between 500 and 2 000 base pairs. RACE reactions were done with the adaptor primers and the gene-speeifie primers designed according to one EST sequence from silk grand in Argiope amoena. By cloning and sequencing of amplified products, the full-length eDNA sequence of the gene which the EST belongs to was obtained. The library was appropriate for cloning the full-length cDNAs of these genes expressed in the silk gland of Argiope amoena. In addition, the features and limitations of SMART technique were discussed.