【摘要】 目的 :将人重组成纤维细胞生长因子受体 1(FGFR1)表达在昆虫细胞膜表面 ,用作筛选成纤维细胞生长因子(FGFs)拮抗肽探针。方法 :将人FGFR1cDNA克隆入昆虫病毒的转递质粒pFastBacI上 ,然后转座到昆虫病毒Bacmid上。以重组Bacmid转染昆虫细胞Sf9并表达人FGFR1,以Western印迹和ELISA对表达出的蛋白质进行鉴定。结果 :FGFR1cDNA片段2 10 0bp ,重组FGFR1表达产物分子量 78kD。ELISA结果显示 ,人重组FGFR1高效地在昆虫细胞Sf9膜表达。结论 :这个表达系统能很好地表达出人重组FGFR1,并能准确地将其定位到昆虫细胞膜的表面。更多还原

【Abstract】 Objective:Express the recombinant human FGFR1 in order to screen the FGFs.Methods:A cDNA fragment encoding human fibroblast growth factor receptor 1(FGFR1) was isolated by RT PCR from human lung fibroblasts,and then cloned into pCR TM II plasmid and pFastBac1 donor plasmid.Through transposition,recombinant bacmid FGFR1 DNA was formed and was then transfected into Sf9 insect cells to produce recombinant Baculovirus.Sf9 insect cell were infected with recombinant Baculovirus.The harvested culture supernatant was subjected to Western Blot and ELISA analysis.Results:The size of FGFR1 cDNA fragment is 2 100 bp.The MW of expressed recombinant FGFR1 was 78 kD.ELISA showed that human recombinant FGFR1 was expressed on the membrane of insect cell Sf9.Conclusion:The recombinant human FGFR1 can be expressed on the membrane of insect cell Sf9 effectively.更多还原