戊型肝炎病毒ORF1多聚蛋白的不同功能域在细胞内的定位

为了探讨戊型肝炎病毒多聚蛋白ORF1的多个功能域在宿主细胞中的表达和定位情况,我们首先将psk-HEV重组载体上的ORF1各功能域的编码序列克隆到绿色荧光蛋白载体pcDNA3.1-GFP上,构建成融合表达的重组质粒,并测序和酶切鉴定其构建成功。再通过Western-Blot验证各融合蛋白在细胞中正确表达,并用激光扫描共聚焦显微镜观察融合蛋白在细胞内的分布和定位。在Huh7细胞中,RdRp蛋白主要分布于细胞核内,HEL蛋白以囊泡状分布于细胞核周,MET蛋白以颗粒状存在于细胞核和细胞质中,PLP蛋白呈极性分布于细胞核周,X蛋白在细胞核和细胞质中均存在。各融合蛋白在细胞中的不同定位印证了对这些蛋白质的功能预测和体外研究结果,这为进一步研究HEV不同蛋白功能提供了支持

The preparation of ZnSe Qdots in aqueous solution and the effects of photo-induced fluorescence enhancement

在水相中,以巯基乙酸(TgA)为稳定剂制备了具有短波长荧光的znSE量子点.研究了znSE量子点光诱导荧光增敏的机理,并提出通过补加zn2+和TgA以提高光诱导荧光增敏效率以及所得znSE量子点的稳定性这一新思路.研究结果表明,提高补加的zn2+和TgA的量即可增加znSE量子点表面znS壳层的厚度,更好地钝化其表面,从而不仅可显著提高znSE量子点的荧光量子产率(最高可接近15%),而且可大大地提高其表面的抗氧化性和荧光稳定性.ZnSe Qdots with short wavelength-fluorescence were prepared in aqueous solution with thioglycolic acid(TGA) as capping reagent.The mechanism of photo-induced fluorescence enhancement was studied and it was proposed a novel approach involved in the addition of compensatory Zn2+ ions and TGA to the original ZnSe solution to increase the efficiency of photo-induced fluorescence enhancement and stability of ZnSe Qdots.The research indicated that the thickness of ZnS shell increases with the increase of the quantity of Zn2+ ions and TGA.The ZnS shell made a better passivation of the ZnSe Qdot surface,thus resulted in not only higher fluorescence quantum yield(maximal 15%),but also higher stability of ZnSe Qdots.It was expected that the proposed approach would also provide a novel route to increase the fluorescence quantum yield and stability of other types of Qdot prepared in aqueous solution.浙江省自然科学基金资助项目(Y4080518