CDNA cloning and expression of autoantigen B0-E2

AIM: To express BO-E2 recombinant fusion protein in Escherichia coli for use in early diagnosis of primary biliary cirrhosis (PBC). METHODS: CDNA encoding BCOADC-E2 and OGDC-E2 were obtained by reverse transcription pdymerase chain reaction (RT-PCR), confirmed by DNA sequencing, subcloned non-directionally into the bacterial expression plasmid pET28a(+), and then transformed into E. coli BL21(DE3) to express the recombinant fusion protein, which was identified by SDS-PAGE and western blotting. RESULTS: Results of nucleotide sequencing and restriction analysis showed that the recombinant plasmid successfully constructed three species. Three types of fusion protein were expressed by isopropyl-Beta-d-thiogalactopyranoside (IPTG) induction. The recombinant fusion protein exhibited the antigenic-ity of AMA-M2 by western blotting. CONCLUSION: BO-E2 recombinant fusion protein can be used for in vitro diagnosis of PBC.link_to_subscribed_fulltex