Construction and expression of cell-penetrating peptide CCL fusion protein expression vector / 中国感染控制杂志

ABSTRACT

Objective To evaluate the construction of expression vector for fusion protein of cell-penetrating pep-tide CCL (PEP-CCL).Methods CCL6-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 chromatog-raphy and detected with Western Blot.Results PEP-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.Conclusion PEP-CCL express vector can be constructed and expressed in eukaryotic cells.