title: [Expression of the antibacterial peptide CM4-like gene of Chinese silkworm Bombyx mori in Escherichia coli and its antibacterial activity analysis].
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.
authors.affiliation: The Life Science School of Nanjing Normal University, Nanjing 210097. jslbc@126.com
authors.forename: Bao Cun
authors.initials: BC
authors.lastname: Li
authors.forename: Yu Qing
authors.initials: YQ
authors.lastname: Chen
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
authors.forename: Shuang Quan
authors.initials: SQ
authors.lastname: Zhang
mesh_terms: 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
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; C534224:CM4 peptide, Bombyx mori; D019476:Insect Proteins; D011993:Recombinant Fusion Proteins; D007544:Isopropyl Thiogalactoside