Construction and expression of fused vector of PAcA and CTB

Objective:To constructe the expression vector of fused gene that the encoding gene of PAcA were linked with the encoding gene of Cholera Toxin B subunit by five peptide, and express the fussion protein. Methods:The pacA ActxB1 amplified by PCR. The DNA products of pacA ActxB1were inserted the prokaryotic expression vector pET32a(+),and then transformed into E.coli strain BL-21(DE3) to express the fusion protein after the induction by IPTG. The recombinant plasmid named pEAC5 was identified by restriction endonuclease analysis, PCR digesion and DNA sequencing. the fusion protein was analysised by SDS-PAGE. Results:The phase and the orientation of the target genes and the linkers which inserted into the pET32a(+) were correct and their ORFs didn't change. the molecular weight of the fusion protein was concerted with the prediction . Conclusions: the recombinant plasmid (pEAC5) was constructed and expressed correctly.