Construction and expression of cellpenetrating peptide CCL fusion protein expression vector

Abstract:

ObjectiveTo evaluate the construction of expression vector for fusion protein of cellpenetrating peptide CCL (PEPCCL). MethodsCCL6PEP6XHis was inserted into plasmid pABP, pABPCCL6PEP plasmid was extracted and then transfected into HEK293 cells, CCL6PEP6XHis was expressed and purified by chromatography and detected with Western Blot. ResultsPEPCCL express vector was successfully constructed and purified. PCR product of CCL6PEP6XHis Tag was ligated with T vector, recombinant was transferred into the host cells, then host cells were cultured, plasmid was extracted and sequenced, the sequence was identical to targeted gene. CCL6PEP6XHis was successfully inserted into the eukaryotic expression vector pABP, plasmid was extracted and digested, electrophoresis results revealed that a fragment with 430bp was digested by Hind Ⅲ+XbaⅠ, which was identical to the expected value. Western Blot revealed that CCL6PEP fusion protein could be recognized by His monoclonal antibody. ConclusionPEPCCL express vector can be constructed and expressed in eukaryotic cells.