紫苏DGAT1基因克隆及四尾栅藻表达载体构建

生物技术通报 ›› 2014, Vol. 0 ›› Issue (5): 137-141.

紫苏DGAT1基因克隆及四尾栅藻表达载体构建

王莹莹¹ 刘玉² 姬妍茹² 张正海² 周广麒¹ 刘宇峰²

(1.大连工业大学生物工程学院，大连 116034；2.黑龙江省科学院大庆分院，大庆 163319)

收稿日期:2013-12-09 出版日期:2014-05-23 发布日期:2014-05-24
作者简介:王莹莹，女，硕士研究生，研究方向：微藻生物技术；E-mail：497035670@qq.com
基金资助:
大庆市科技计划项目(scyh-2011-76)

Cloning of DGAT1 Gene from Perilla frutescens and Construction of Scenedesmus quadricanda Expression Vector

Wang Yingying¹ Liu Yu² Ji Yanru² Zhang Zhenghai² Zhou Guangqi¹ Liu Yufeng²

(1. School of Bioengineering, Dalian Polytechnic University, Dalian 116034；2. Daqing Branch of Heilongjiang Academy of Sciences, Daqing 163319)

Received:2013-12-09 Published:2014-05-23 Online:2014-05-24

摘要/Abstract

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。

关键词: 四尾栅藻, DGAT1基因, 克隆, 表达载体构建

Abstract: Total RNA of Perilla frutescens was extracted by RNAiso Plus kit. It was used as a template for RT-PCR to clone cDNA coding region of DGAT1 gene of Perilla frutescens, and cloning vector pMD-DGAT1 was constructed. Cloning vector pMD-DGAT1 and empty vector pBI121 were double-enzyme digested with Xba I and BamH I. Then expression vector pBI121-DGAT1 was constructed by linking the fragments. Results showed that 1 657 bp fragment was obtained by cloning and has 97% base similarity to DGAT1 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 Scenedesmus quadricanda successfully. The transformed Scenedesmus quadricanda approximately double the oil content as before.

Key words: Scenedesmus quadricanda, DGAT1 Gene, Clone Expression vector, Construct

王莹莹, 刘玉, 姬妍茹, 张正海, 周广麒, 刘宇峰. 紫苏DGAT1基因克隆及四尾栅藻表达载体构建[J]. 生物技术通报, 2014, 0(5): 137-141.

Wang Yingying, Liu Yu, Ji Yanru, Zhang Zhenghai, Zhou Guangqi, Liu Yufeng. Cloning of DGAT1 Gene from Perilla frutescens and Construction of Scenedesmus quadricanda Expression Vector[J]. Biotechnology Bulletin, 2014, 0(5): 137-141.

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