The Construction and Expression of A Fusion Protein Consisting of Anti-HBsAg Single Chain Fv and lnterleukin-2 E.coli

Objective: To construct an expression vector for anti-HBsAg ScFv and interleukin-2 fusion protein in E. coli-2. Methods: Using DNA recombination and overlap PCR technology, a cloned vector PGEM7Zf( + )/ScFv for anti-HBsAg ScFvand interleukin-2 fusion protein was constructed. Enzyme digestion and DNA sequence measurement confirmed that the ScFv-IL-2 gene had been correctly cloned to the vector, and then the target gene was cloned to the expression vector PQE40, choosing the passive clone to transform into E. coli and induced by IPTG to expression protein. Results: A recombinant expression plasmid PQE40/ ScFv-IL-2 was constructed. 4. 3 kU expression protein was observed by SDS-PAGE, and Western blot with mouse anti-human IL-2 McAb showed that the expressed products were designed fusion protein. Conclusion: The success in construction and expression of fusion protein makes it possible to carry out further studies on its purification and the targeted therapy of gene-engineering pharmaceutic to HB virus.