Gene Cloning,Expression and Purification of sRNA Chaperone hfq of Listeria monocytogenes

doi:10.7668/hbnxb.2014.05.014

Abstract

Abstract: In order to clone,express the Listeria monocytogenes sRNA chaprone hfq gene and purify the recombinant protein,the hfq gene was amplified by PCR from LM genome,the amplified products was cloned in pMD18-T vector,sequenced and subcloned in the expression vetor pET-32a(+).The recombinant pET-32a(+)-hfq expression vector was generated successfully and then was transformed into E.coli competent cells of BL21(DE3),induced by IPTG.The solubility of Hfq protein was analyzed and then the recombinant protein was purified by the Ni-NTA affinity chromatography.The result showed the hfq gene had a length of 234 bp,encoding 77 amino acids.The specific 27 kDa fusion protein was examined by SDS-PAGE.Finally,the fusion protein was obtained successfully from the supernatant.This study laid the foundation for further study the biological function of the Hfq protein.

Key words: Listeria monocytogenes, hfq gene, sRNA chaperone, Cloning, Expression, Purification

CLC Number:

S852
Q78

PENG Ye-long, QIAO Jun, MENG Qing-ling, XIE Kun, CHEN Cheng, LIU Tian-li, MA Yu, CAI Xue-peng, CHEN Chuang-fu. Gene Cloning,Expression and Purification of sRNA Chaperone hfq of Listeria monocytogenes[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(5): 80-87. doi: 10.7668/hbnxb.2014.05.014.

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