摘要
利用RNAiso Plus试剂盒提取紫苏总RNA,以其为模板采用RT-PCR方法扩增出紫苏DGAT1基因的cDNA编码区序列并构建克隆载体pMD-DGAT1,再将克隆载体pMD-DGAT1和pBI121空载体进行Xba I和BamH I双酶切,连接目的片段构建表达载体pBI121-DGAT1。结果显示,克隆得到目的片段长度为1 657 bp,与GenBank中紫苏DGAT1基因序列的最大同源性为97%,所构建表达载体pBI121-DGAT1双酶切得到的两条片段长度与连接前一致。结果表明,成功转化四尾栅藻并得到表达,转化后四尾栅藻的油脂含量较转化前提高近1倍。
Total RNA of Perillafrutescens was extracted by RNAiso Plus kit. It was used as a template for RT-PCR to clone cDNA coding region of DGAT1 gene of Perillafrutescens, and cloning vector pMD-DGAT! was constructed. Cloning vector pMD-DGAT1 and empty vector pBI 121 were double-enzyme digested with Xba I and BamH I. Then expression vector pBI 12 1-DGAT 1 was constructed by linking the fragments. Results showed that 1 657 bp fragment was obtained by cloning and has 97% base similarity to DGATI gene of GenBank. Expression vector pBI121-DGAT1 was double-enzyme digested into two fragments. The length of them was as long as the ones before linking. It was transformed and expressed in the Scenedesmu,~ quadricanda successfully. The transformed Scenedesmus quadricanda approximately double the oil content as before.
出处
《生物技术通报》
CAS
CSCD
北大核心
2014年第5期137-141,共5页
Biotechnology Bulletin
基金
大庆市科技计划项目(scyh-2011-76)
关键词
四尾栅藻
DGAT1基因
克隆
表达载体构建
Scenedesmus quadricanda DGAT1 gene Clone Expression vector construct
