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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 HIV-1 envelope protein gp41 was displayed on the surface of MT8-1 of Saccharomycis cerevisiase using Hexa-His and Flag as a tag, and the results of flow cytometry showed that His tag was successfully expressed in recombinant yeast cells MT8-1 / pICAS-His and MT8-1 / pICAS-His-gp41. The active gp41 was successfully displayed on the surface of recombinant yeast MT8-1 / pICAS-His-gp41. The results of fluorescence microscopy showed this point. Detected on the surface of TM8-1 / pICAS-Flag, neither Flag nor gp41 detected activity on the surface of MT8-1 / pICAS-Flag-gp41.Using gp41 protein displayed on the surface of yeast to establish yeast cell CbELISA system can be successfully used For the detection of monoclonal antibodies at a concentration of 10 ng / mL, this system can be used to detect antibodies in serum.