Prokaryotic expression and purification of PEP-1-VP3 fusion protein

OBJECTIVE To construct protaryotic expression vector p ET15b- PEP- 1- VP3 induce expression and to purify the fusion protein of PEP- 1- VP3. METHODS To VP3 c DNA was amplified from the plasmid of PGEX- 6P- 1 / TAT- VP3 by PCR.Then the amplified VP3 c DNA was cloned into linearized p ET15b- PEP- 1 by LP Recco PCR cloning. The recombinant plasmid p ET15b- PEP- 1- VP3 were identified by sequencing analysis and transformed into E. coli Rosetta for expression of PEP- 1- VP3 fusion protein. After purified by Ni2 +- resin affinity chromatography,expressed products were identified by SDS- PAGE and Western Blot. RESULTS The recombinant plasmids were constructed and PEP- 1- VP3 fusion protein with relative molecular weights of 17. 6k D was expressed successfully. CONCLUSION The PEP- 1- VP3 fusion protein could provide a basis using in the inducing apoptosis of tumor cells.