Cloning and Expression of HBV Infection Related Novel Gene C12orf49

摘要

Objective To clone, express and purify C12orf49 recombinant protein. To prepare rabbit anti-C12orf49 protein polyclonal antibody and further elucidate its biological function. Methods PCR was applied to amplify the gene C12orf49 in vitro. pET-32a (+)-C12orf49, the recombinant protein prokaryotic expression vector, was transformed into E. coli. IPTG was used as an inductive agent to obtain C12orf49 recombinant protein, then the recombinant protein was analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Specific polyclonal antibody was derived from rabbits that was immunized by recombinant protein. ELISA and Western blot were used to detect its titer and speciifcity, respectively. MTT cell proliferation experiment was carried out to observe the effect of protein on proliferation of HepG2 cells. Results The C12orf49 recombinant protein was expressed in a large quantity. Data of ELISA indicated that the titer of polyclonal antibody was higher than 1:1 280 000. And a good speciifcity of the antibody was conifrmed by Western blot. C12orf49 recombinant protein might have an advanced effect on the proliferation of HepG2 cells. Conclusions Using C12orf49 recombinant protein, we can obtain the polyclonal antibody with great titer and good speciifcity. Human novel gene C12orf49 encoded protein could promote the proliferation of HepG2 cells.