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Fen Zi Xi Bao Sheng Wu Xue Bao. 2007 Apr;40(2):98-102.

[Expression of the antibacterial peptide CM4-like gene of Chinese silkworm Bombyx mori in Escherichia coli and its antibacterial activity analysis].

[Article in Chinese]

Author information

1
The Life Science School of Nanjing Normal University, Nanjing 210097. jslbc@126.com

Abstract

According to the amino acid sequence of CM4 and the bias for preferred condons of E. coli, the CM4-like gene was obtained by a recursive PCR (rPCR) strategy using two lapping oligonucleotides. The synthesized gene was coloned into the expression vector pET32(a) and transformed into E. coli BL21 (DE3). Recombinant CM4-like gene expression was driven by the T7 promoter on the vector upon addition of IPTG and high level of expression was achieved. The solube protein was purified by Ni-chelating agarose and treated with formic acid. After cleavege, the recombinant peptide was purified by another Ni(2+)-NTA-Agarose affinity chromatography and cation-exchange chromatography. Results of agarose diffuse assay and liquid turbidity analysis indicated that the recombinant peptide exhibited the antibacterial activity.

PMID:
17580662
[Indexed for MEDLINE]

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