Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (5): 137-141.

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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. (1. School of Bioengineering, Dalian Polytechnic University, Dalian 116034;2. Daqing Branch of Heilongjiang Academy of Sciences, Daqing 163319)
  • Received:2013-12-09 Online:2014-05-23 Published:2014-05-24

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