[Fusion of betal-toxin gene and beta2-toxin gene from Clostridium perfringens type C].
Betal-toxin and beta2-toxin genes from chromosomal DNA of Clostridium perfringens type C were amplified by PCR, PCR products were cleaved with restriction endonucleases and recovered. The recombinant plasmid pETXB1-2 containing beta1-beta2 fusion genes was constructed by recombinant technique and then transformed into Escherichia coli BL21 (DE3). The beta1-beta2 fusion proteins were expressed in recombinant strain BL21 (DE3) (pETXB1-2), and the expression level of the beta1-beta2 fusion proteins was about 15.36% of total cellular protein by SDS-PAGE and thin-layer gel scanning analysis. More importantly, immunization in a mouse model with crude preparation containing the fusion protein inclusion bodies or inactivated recombinant strain induced protection against at least 1 MLD of the toxin from Clostridium perfringens type C. Hence the fusion proteins possess a good immunogenicity. The constructed recombinant strain BL21 (DE3) (pETXB1-2) can be used as a candidate of vaccine strain.
College of Bioengineering, Dalian University, Dalian 116622, China. xcb921@sohu.com
Chong-bo
CB
Xu
Jin
J
Zeng
Chong-li
CL
Xu
Yu-jiong
YJ
Wang
D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics* / Q000276:immunology; D003016:Clostridium perfringens / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D050939:Gene Fusion; D005798:Genes, Bacterial; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology
D001427:Bacterial Toxins; C031884:CPB protein, Clostridium perfringens; D011993:Recombinant Fusion Proteins; C109896:cpb2 protein, Clostridium perfringens