【摘要】 目的 :构建TGF βRⅡ /Fc融合蛋白的杆状病毒表达系统 ,并使之成功表达 .为将来TGF βRⅡ /Fc融合蛋白在肺间质纤维化基因治疗中的应用奠定基础 .方法 :采用分子克隆技术构建Bac TRⅡ杆状病毒表达系统 ;采用DNA脂质体转染技术和昆虫细胞培养技术培养和扩增Bac TRⅡ重组病毒颗粒 ;采用免疫荧光技术和免疫印迹法检测目的蛋白的表达 .结果 :在重组的Bac TRⅡ杆状病毒感染的sf9细胞中可以检测到目的蛋白的表达 .结论 :重组的Bac TRⅡ杆状病毒表达系统可以用于正确表达TGF βRⅡ /Fc融合蛋白 .由于杆状病毒表达系统效率较高 ,因此 ,今后可利用该系统进行目的蛋白的纯化及进一步的实验研究 .更多还原

【Abstract】 AIM: To construct the baculovirus expression system of chimeric protein TGF βRⅡ/Fc, and have it successfully expressed for future application of chimeric protein TGF βRⅡ/Fc in gene therapy of pulmonary fibrosis. METHODS: Recombinant Bac TRⅡ baculovirus expression system was constructed by molecular cloning technology; The DNA liposome transfection technology and insect cell culture technology were employed to culture and amplify the recombinant Bac TRⅡ baculovirus particles. The chimeric protein was detected by cellular immunofluorescence technique and Western blotting. RESULTS: The chimeric protein was detected in sf9 cells infected by recombinant Bac TRⅡ baculovirus particles. CONCLUSION: The recombinant Bac TRⅡ baculovirus expression system can be correctly used to express chimeric protein TGF βRⅡ/Fc. The baculovirus expression system can efficiently express the target protein and facilitate the purification of the target protein.更多还原