P35识别结合HCV糖蛋白的研究

Research of P35 About Its Binding to HCV E1

目的:研究P35(L-ficolin)作为一种凝集素补体,能否结合HCV包膜糖蛋白E1。方法:用PC DNA3-P35质粒转染细胞,建立稳定表达P35的细胞系CT26-P35,RT-PCR和Western Blot检测该细胞系,建立稳定表达HCV E1蛋白的细胞系E1-CT26,用FCM检测该细胞胞内胞膜上E1的表达。用FCM检测蛋白P35与HCV E1的作用。结果:成功建立稳定细胞系P35-CT26。E1-CT26细胞胞内和胞膜均有E1表达。FCM分析,P35与E1-CT26结合能力强于P35与CT26的结合。结论:成功建立稳定表达P35的细胞系;P35能识别并结合HCV的糖蛋白E1。

Objective： To study whether P35 can bind to HCV E1 as a kind of C-type lectin. Methods.. The vector PCDNA3-P35 was constructed and transfect it into CT26 cells. RT-PCR and Western Blot were used to identify the P35-CT26 cell line. And a stable cell line, E1-CT26, was constructed before the E1 and the P35＇s binding ability to HCV E1 was determined by FCM. Results： The stable cell line P35-CT26 had been successfully constructed. HCV E1 could express both in the cytoplasm and on the surface of E1-CT26. FCM revealed that P35 could bind to HCV E1 more tightly than CT26 does. Conclusion： The stable cell line P35-CT26 had been successfully constructed and P35 could recognize and bind to HCV El.