Prokaryotic Expression Of PET28a(+)-SY1 Phosphatase Recombinant And Primary Preparation Of Its Monoclonal Antibody

Objective: To construct and express the recombination vector of pET28a(+)-SY1 plasmid (constructed by inserting the full-size human SY1cDNA) and to make monoclonal antibody for SY1 to identify basic biologic activity of it. Method: We amplified the SY1 by PCR, digested it by double restriction endonuclease, recombined it into pET28a(+), then transfected into BL21(DE3), added IPTG to induce its expression and detected it by SDS-PAGE and Western Blot. The recombinant SY1 protein was purified on Ni2+-NTA column uNher denature condition. Then the purified fusion protein of SY1 was inoculated in the BALB/c mice as antigens to prepare monoclonal antibody. Indirect ELISA was used to detect the titer. Results: The recombination plasmid was constructed and the fusion protein was induced by IPTG, SDS-PAGE analysis showed the expression protein was mainly in the form of inclusion. The monoclonal antibodies anti SY1 protein were acquired after cell fusion and coloning. Conclusions: We construct and express the vector pET28a(+)-SY1 successfully and obtain the monoclonal antibody(McAb) with the fusion protein as antigen.The successful development of anti-SY1 antibody provides a powerful tool for further investigation on SY1 's function.