Koroleva N N, Spirin P V, Timokhova A V, Rubtsov P M, Kochetkov S N, Prasolov V S, Belzhelarskaia S N
Mol Biol (Mosk). 2010 May-Jun;44(3):541-50.
Baculovirus expression vectors are extensively used for the delivering foreign genes and expression of recombinant proteins in insect and mammalian cells. Modified baculoviruses containing mammalian promoter elements (BacMam viruses) for an efficient transient and stable transduction of diverse mammalian cells prove a high level of heterologous proteins' expression both in vitro and in vivo. Recombinant baculovirus vectors containing mammalian expression cassette with cytomegalovirus promoter, green or red fluorescent protein gene, polyadenylate signal SV40pA, and polylinker MCS were constructed for the delivery of genes encoding hepatitis C virus structural proteins into mammalian cells. In Hek293T and Huh7 cells formation of glycoprotein complexes and HCV-like particles was observed. A high efficiency of the baculovirus-mediated gene transfer and expression of the virus envelope proteins in mammalian cells was demonstrated with using fluorescence, flow cytometry and immunoblot techniques.
杆状病毒表达载体被广泛用于在昆虫和哺乳动物细胞中递送外源基因和表达重组蛋白。含有哺乳动物启动子元件的修饰杆状病毒(BacMam病毒)可有效瞬时和稳定转导多种哺乳动物细胞,在体外和体内均能实现高水平的异源蛋白表达。构建了含有带有巨细胞病毒启动子、绿色或红色荧光蛋白基因、聚腺苷酸信号SV40pA和多克隆位点MCS的哺乳动物表达盒的重组杆状病毒载体,用于将编码丙型肝炎病毒结构蛋白的基因递送至哺乳动物细胞。在Hek293T和Huh7细胞中观察到了糖蛋白复合物和丙肝病毒样颗粒的形成。利用荧光、流式细胞术和免疫印迹技术证明了杆状病毒介导的基因转移在哺乳动物细胞中的高效性以及病毒包膜蛋白的表达。