Journal of South China University of Technology (Natural Science Edition) ›› 2008, Vol. 36 ›› Issue (4): 122-126.

• Biology Science • Previous Articles     Next Articles

Cloning and Fusion Expression of Gene MalQ from E. coil

Wang Shui-xing1  Guo Yong1  Xu Yang2  Li Yan-ping2   

  1. 1. School of Biological Science and Engineering, South China University of Technology, Guangzhou, 510006, Guangdong, China; 2. Jiangxi-OAI Research Institute, Nanchang University, Nanchang, 330047, Jiangxi, China
  • Received:2007-03-07 Revised:2007-04-19 Online:2008-04-25 Published:2008-04-25
  • Contact: 郭勇，教授，博士生导师． E-mail:feyguo@scut．edu．cn
  • About author:王水兴（1965-），男，博士，南昌大学副教授，主要从事食品生物技术研究．E-mail：shuixingw@yahoo．com．cn.
  • Supported by:

    广东省重点科技攻关项目（A301020201）

PDF

921

Abstract:

In order to construct a gene engineering bacterium expressing amylomaltase, gene MalQ was amplified from Escherichia coli K12 by PCR, and was cloned into pET-DsbA. Then, a two-direction sequencing was performed, and the results were analyzed by means of BLAST. There displayed high similarity （99%） of the obtained gene MalQ to the gene MalQ sequence of E. coli K12 reported in the GenBank. Moreover, the recombined plasmid pET-DsbA-MalQ was transformated into E. coli BL21 （DE3） plysS and was induced by IPTG. The fusion protein of gene MalQ and DsbA with a relative molecular mass of about 103000 was detected by means of sodium dodecyl sul- phate-polyacrylamide gel electrophoresis. 4-α-glucanotransferase activity of the crude enzyme was finally demonstrated by means of thin layer chromatography.

Key words: amylomahase, gene recombination, fusion expression, cycloamylose, gene clone