[Expression of the antibacterial peptide CM4-like gene of Chinese silkworm Bombyx mori in Escherichia coli and its antibacterial activity analysis].
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.
Li
Bao Cun
BC
The Life Science School of Nanjing Normal University, Nanjing 210097. jslbc@126.com
Chen
Yu Qing
YQ
Liu
Ping
P
Zhang
Shuang Quan
SQ
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism*; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D047108:Embryonic Development / Q000502:physiology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; C534224:CM4 peptide, Bombyx mori; D019476:Insect Proteins; D011993:Recombinant Fusion Proteins; D007544:Isopropyl Thiogalactoside