Journal of South China University of Technology (Natural Science Edition) ›› 2007, Vol. 35 ›› Issue (11): 110-114.

• Biotechnology • Previous Articles     Next Articles

Cloning and Expression of Conjugated Linoleic Acid Isomerase Gene of Lactobacillus acidophilus

河南工业大学 生物工程学院，闽南 郑州 450052   

  1. College of Biological Engineering， Henan Unive. of Tech. , Zhengzhou 450052 ， Henan , China
  • Received:2006-11-08 Online:2007-11-25 Published:2007-11-25
  • Contact: 曹健(1969-) ，女，博士，教授，主要从事食品微生物学研究. E-mail:biotech@haut.edu.cn
  • About author:曹健(1969-) ，女，博士，教授，主要从事食品微生物学研究.
  • Supported by:

    河南省杰出青年科学慕金资助项目(04120001800 ;问南省高校杰出科研人才创新工程项目(2006KYCX008) ;湾南省教育厅高等学校部新人才培养工程(豫教高[2005) 126 号)资助项目

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Abstract:

This paper deals with the cloning and expression of conjugated linoleic acid isomerase gene of Lactobaciι lus αcidophilus AS 1. 1854 by means of molecular biologïcal method. In the investigation , first , the genome was ex-tracted after the cel1 was cultured and gathered , and was amplified by means of PCR. Next , the amplified gene was cloned into pET30a plasmid vector and subsequently converted into Escherichia coli BL21 strain. Then , the gene was induced to express the recombinant protein at a low temperature , and the recombinant protein was separated and purified with Ni-NTA agarose. Finally , the target gene was amplified and converted into plasmid , and the re-combinant protein was expressed and separated through the experiment. It is found that soluble recombinant conju-gated linoleic acid isomerase can be expressed in prokaryotic cell, and that the target protein can be obtained by washing wÎth 200mmol/L imidazole.

Key words: Lactobacillus acidophilus, conjugated linoleic acid isomerase, gene cloning, recombinant protein, pu-rification