Constrction and prokaryotic expression of a new anti-HIV targeted toxin
抗AIDS新型导向毒素CVN-LP1融合基因的构建及原核表达

Objective: To construct prokaryotic expression vector containing CVN-LP1 gene ,and identify the expression of CVN-LP1 protein. Methods: pET-CVN and pET-LP1 was restriction endonuclease digested by EcoRI and HindIII, and the CVN gene was inserted into pET-LP1 plasmid. The vector was transformed into BL21(DE3) to construct a prokaryotic expression system. The expressed product was identified by SDS-PAGE and Western blot assay. Results: The recombinant plasmid pET-CVN-LP1 was constructed, restriction endonuclease digestion and PCR identification proved that the CVN was correctly cloned into vector pET-LP1. SDS-PAGE and Western-blot analysis showed that CVN-LP1 was successfully expressed in E.coli BL21(DE3) ,The relative molecular mass (Mr) of the expression protein was 20kDa,according with the predicted Mr value. And the expression yield was about 49.58% of total bacterial protein. Conclusion: The successful expression of CVN-LP1 gene in E.coli provides a support for further study of the anti-HIV agent.