Expression in E. coli, Purification and Refolding of Recombinant Cry1C Gene from Bacillus thuringiensis

doi:10.7506/spkx1002-6630-201107045

FOOD SCIENCE ›› 2011, Vol. 32 ›› Issue (7 ): 211-215.doi: 10.7506/spkx1002-6630-201107045

• Bioengineering • Previous Articles Next Articles

Expression in E. coli, Purification and Refolding of Recombinant Cry1C Gene from Bacillus thuringiensis

CAO Si-shuo1,XU Wen-tao1,2,RAN Wen-jun1,LIANG Li-xing1,HE Xiao-yun2,LUO Yun-bo1, YUAN Yan-fang2,HUANG Kun-lun1,2,

1.College of Food Science and Nutrition Engineering,China Agricultural University,Beijing 100083,China; 2.Supervision,Inspection and Testing Center of Genetically Modified Food Safety,Ministry of Agriculture,Beijing 100083,China

Received:2010-06-03 Revised:2011-02-25 Online:2011-04-15 Published:2011-03-30
Contact: XU Wen-TAO E-mail:xuwentao1111@sina.com

Abstract

Abstract: The Cry1C gene from Bacillus thuringiensis (Bt) was cloned by polymerase chain reaction (PCR). Due to a large number of E. coli low-usage codons in the gene, the first 86 bases were optimized by PCR to improve the expression in E. coli. Cry1C protein was highly expressed in E. coli BL21 as inclusion bodies, which can be dissolved in 8 mol/L urea and purified by His TrapTM FF crude column under denaturing conditions. The purified Cry1C protein was dialyzed against the refolding buffer to obtain a soluble and biologically active protein. Finally, the protein was determined to be 99.2% purity.

Key words: Cry1C protein, rare codon, inclusion body, expression, purification, refolding

CLC Number:

Q344.13

CAO Si-shuo,XU Wen-tao,RAN Wen-jun,LIANG Li-xing,HE Xiao-yun,LUO Yun-bo, YUAN Yan-fang,HUANG Kun-lun1. Expression in E. coli, Purification and Refolding of Recombinant Cry1C Gene from Bacillus thuringiensis[J]. FOOD SCIENCE, 2011, 32(7 ): 211-215.

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Expression in E. coli, Purification and Refolding of Recombinant Cry1C Gene from Bacillus thuringiensis

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