酵母细胞表面展示HIV-1 gp41用于血清中抗体的检测的研究(英文)  

STUDIES ON THE HIV-1 GP41 DISPLAY ON YEAST SURFACE FOR ANTIBODY DETECTION IN SERA

作　　者：向柱方[1] 赵树进[2] 杜红丽[1] 林影[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006 [2]广州军区总医院,广东广州510010

出　　处：《陕西科技大学学报（自然科学版）》2008年第2期16-23,共8页Journal of Shaanxi University of Science & Technology

基　　金：广东省科技厅专项基金(No.G02B2041120)

摘　　要：以Hexa-His和Flag为标签,利用酵母表面展示技术将HIV-1囊膜蛋白gp41展示于酿酒酵母(Saccharomycws cerevisiase)细胞MT8-1表面.流式细胞测定结果表明His标签成功展示于重组酵母细胞MT8-1/pICAS-His和MT8-1/pICAS-His-gp41表面;活性的gp41成功展于重组酵母MT8-1/pICAS-His-gp41表面,荧光显微镜观测结果表明了这一点;Flag标签可以在TM8-1/pICAS-Flag表面检测到,但Flag和gp41均不能在MT8-1/pICAS-Flag-gp41表面检测到活性.利用展示在酵母表面的gp41蛋白建立酵母细胞CbELISA体系,可成功用于检测单克隆抗体,其浓度达到了10 ng/mL,此体系可用于检测血清中的抗体.The envelope glycoprotein gp41 of HIV-1 has been successfully displayed on the cell surface of Saccharomyces cerevisiase MT8-1 by cell-surface engineering using Hexa-His and Flag tags for detection, respectively. Flow cytograms showed that His tag was displayed on the surface of MT8-1/pICAS-His and MT8-1/pICAS-His-gp41, the active gp41 could be detected on the cell surface of MT8-1/pICAS-His-gp41 and was confirmed by fluorescence microscopy; Flag tag was detected on the surface of MTS-1/pICAS-Flag, but Flag tag and gp41 were not detected on the surface of MT8-1/pICAS-Flag-gp41 separately. Further, a yeast CbELISA was developed by using the yeast cells displaying gp41 on the cell surface. It was able to detect the monoclonal antibody at the concentration of 10 ng/mL , and also capable of detecting the antibody in the sera.

关 键 词：HIV-1 GP41 酵母细胞CbELISA 免疫荧光标记 FACS(流式细胞检测技术) 

分 类 号：Q786[生物学—分子生物学]