关键词: 穿膜肽, 增强型绿色荧光蛋白, 膀胱肿瘤

Abstract: Objective To construct EGFP-CPPs fusion genetic carrier,express,purify and identify EGFP-CPPs fusion protein.Methods Upstream primer containing CPPs gene sequence of green fluorescent was synthesized by basic radical synthesise methods.CPPs-EGFP fusion gene fragment was amplificatied by PCR methods and was indentified by electrophoresis. Bacillus coli were transfected with the fusion gene express vector and the fusion gene was amplificated and expressed.Fusion protein was collected and purified by column chromatography.Protein molecular mass was detected by protein electrophoresis.Protein sequence was detected by TOF analysis.Cell-penetrating function was tested by vitro cell experiment. Results Electrophoresis site corresponding to EGFP-CPPs was displayed in electrophoresis;The gene sequence of constructed plasmid contained interest gene sequence.Purified fusion protein was displayed in protein electrophoresis.Protein sequence was in accord with interested protein sequence. CPPs-EGFP fusion protein displayed Cell-penetrating function in vitro. Conclusions CPPs-EGFP fusion protein with cell-penetrating function was prepared successfully in the experiment.It will establish basis for study of promoting antibody-albumin nanosphere conjugate to enter tumor cell and bring into full play therapeutical effect.

Key words: CPPs, EGFP, Bladder tumor