CAJ | 학술논문

目的分离和初步鉴定口本脑炎病毒(JEV)易感细胞C6/36和Vere细胞上受体分子.方法应用免疫共沉淀(Co-IP)技术,从C6/36和Vero细胞分离与JEV结合的受体分子,做质谱分析鉴定和Western blot检测.用激光共聚焦技术(LSCM)观察候选受体分子在细胞膜上的定位及与JEV结合的情况.结果经过Co-IP反应,从C6/36、Vem细胞上分离出多个与JEV结合的分子条带.质谱分析首次鉴定出一个蛋白,即C6/36细胞上与JEV结合的相对分子质最(Mr)为74×103分子,为HSC70蛋白.进一步用抗HSC70抗体做Western blot,榆测剑从C6/36和Vem细胞膜上Co-IP分离出的74×103蛋白.LSCM观察到JEV吸附在C6/36细胞膜上时,HSCT0蛋白与JEV共定位.结论 C6/36细胞上Mr为74×103的HSC70可能是JEV的受体分子.
목적 분리화초보감정구본뇌염병독(JEV)역감세포C6/36화Vere세포상수체분자.방법 응용면역공침정(Co-IP)기술,종C6/36화Vero세포분리여JEV결합적수체분자,주질보분석감정화Western blot검측.용격광공취초기술(LSCM)관찰후선수체분자재세포막상적정위급여JEV결합적정황.결과 경과Co-IP반응,종C6/36、Vem세포상분리출다개여JEV결합적분자조대.질보분석수차감정출일개단백,즉C6/36세포상여JEV결합적상대분자질최(Mr)위74×103분자,위HSC70단백.진일보용항HSC70항체주Western blot,유측검종C6/36화Vem세포막상Co-IP분리출적74×103단백.LSCM관찰도JEV흡부재C6/36세포막상시,HSCT0단백여JEV공정위.결론 C6/36세포상Mr위74×103적HSC70가능시JEV적수체분자.
Objective To isolate and identify the putative Japanese encephalitis virus (JEV) re-ceptors from C6/36 and Vero cells. Methods Molecules binding with JEV were isolated from C6/36 and Vero cells by co-immunoprecipitation (Co-IP) approach, identified by mass spectrometry, and detected by Western blot. The location of putative JEV receptor on cells membrane and the binding with JEV were ob-served by laser scanning confocal microscopy (LCM). Results Several molecules binding with JEV were isolated from C6/36 and Vero cells by Co-IP, and only one molecule was identified as heat shock cognate 70 (HSC70) by mass spectrometry. Antibody against HSC'70 was able to detect a 74 ×103 protein isolated by Co-IP from C6/36 and Vero cells membrane in Western blot assays. It was observed by LSCM that when JEV attached on the surface of C6/36 cells, JEV and HSCT0 protein were co-localization. Conclusion 74 x 103 molecular identified as HSC70 protein from C6/36 cells may be JEV receptor.