[Expression and purification of an adhesive protein of rabbit Pasteurella multocida C51-3 and detection of its antigenicity].
The cp36 gene encoding an adhesive protein was amplified by PCR from genomic DNA of rabbit P. multocida C51-3 strain, and cloned into the pMD18-T vector and then sequenced. The mature adhesive protein without a signal peptide of cpm36 gene was amplified by PCR from the recombinant plasmid pMD18-cp36, then cloned into the prokaryotic expression vector pQE30 to provide a recombinant plasmid pQE30-cpm36. The recombinant protein of CPM36 was produced in Escherichia coli M15 harboring the recombinant plasmid pQE30-cpm36 by IPTG induction, and the recombinant protein purified by the affinity chromatography with Ni(2+)-NTA resin. The sequence analyses showed that the ORF of cp36 gene was 1032 bp in length, and DNA homology of the cp36 genes between the C51-3 strain and the previously reported different serotype strains of P. multocida in GenBank was 76.9 to 100%. The SDS-PAGE analyses revealed a single fusion protein band with a molecular weight of 37 kD, and the Western blotting analysis demonstrated that the recombinant protein CPM36 and native 36 kD protein of C51-3 were recognized specifically by an antiserum against the recombinant protein, suggesting that the recombinant protein is an antigenic protein.
Nazierbieke
Wulumuhan
W
Laboratory of Bioengineering, College of Biology and Environmental Sciences, Jishou University, Jishou 41600, China.
Yan
Fang
F
He
Cui
C
Zhang
Lei
L
Borrathybay
Entomack
E
D018829:Adhesins, Bacterial / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016979:Pasteurella multocida / Q000737:chemistry*; D011817:Rabbits / Q000382:microbiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D018829:Adhesins, Bacterial; D011993:Recombinant Fusion Proteins