Preparation of Protein Transduction Domain Modified Mice IFN-? / 中国药房

ABSTRACT

OBJECTIVE:To prepare PTD modified mice interferon gamma(PTD-mIFN-?) in order to prepare for its function research.METHODS:The code of PTD-mIFN-? fragment was amplified by PCR for 3 times,using pUC19/mIFN-? as template.The PCR product was cloned into pUC19 plasmid and then constructed into expression plasmid vector pQE80L after sequencing.After double digestion with BamH Ⅰ and Hind Ⅲ,the recombinant vector was identified in 1.2% agarose gel.The target protein and PTD-mIFN-? was checked by SDS-PAGE,Western blot method and ELISA assay,respectively.Recombinant protein was purified by nikel ion-triglycollamic acid(Ni2+-NTA) Agarose affinity chromatogram.RESULTS:The cDNA fragment encoding PTD-mIFN-? amplified by PCR was obtained.The high-level expression of recombinant protein was noted in SDS-PAGE and Western blot assay.The target protein was obtained by ELISA assay.CONCLUSION:PTD-mIFN-? was prepared successfully.