Cloning of α-β fusion gene from Clostridium perfringens and its expression

Figure 1 Identification of α-toxin gene product amplified by PCR (A) and recombinant plasmid by enzyme digestion (B). In A, lanes 1, 15: DNA markers; lanes 2-14: product of α-toxin gene amplified by PCR; In B, lane 1: DNA markers; lanes 2-7: pMD 906/ EcoR I + BamH I; lane 8: pMD 906/ EcoR I.

Figure 2 Nucleotide sequence of α-toxin gene from Clostridium perfringens type A strain NCTC64609.

Figure 3 Identification of β-toxin gene product amplified by PCR (A) and recombinant plasmid by enzyme digestion (B). In A, lanes 1,17: DNA markers; lanes 2-16: product of β-toxin gene amplified by PCR; In B, lane 1: pMD930/EcoR I; lane 2: pMD930/ Pst I; lane 3: DNA Markers; lane 4: pMD930/BamH I+Pst I

Figure 4 Nucleotide sequence of β-toxin gene from Clostridium perfringens type C strain C58-1

Figure 5 Agarose gel electrophoresis of restriction endonucleases-digested plasmid pZCPAB. Lane 1: DNA markers; lane 2: pZCPAB / EcoR I; lanes 3-5: pZCPAB / EcoR I+Pst I; lane 6: pZCPAB / BamH I.

Figure 6 Analysis of expressed products of α-β fusion gene in E. coli by SDS-PAGE (A) and Western blot (B) In A, lane 1: protein markers (97400, 66200, 43000, 31000, 20100, 14400); lanes 2, 3: BL21(DE3)( pZCPAB); lane 4: BL21(DE3)(pBV221); In B, lane 1: BL21(DE3)(pZCPAB)/α; lane 2: BL21(DE3)(pZCPAB)/α+β; lane 3: BL21(DE3)( pZCPAB)/β; lanes 4, 5: BL21(DE3)(pBV221).

Figure 7 Pathological changes in livers of survived mice (A) and dead mice (B), in lung of survived mice (C) and dead mice (D), in heart of survived mice (E) and dead mice (F), in spleen of survived mice (G) and dead mice (H), in intestine of survived mice (I) and dead mice (J).