摘要
The recombinant baculoviruses were constructed to investigate the necessity of VSV-G pseudotyping for mammalian cell transduction. The viruses were designed to express green fluorescent protein (GFP) gene under the control of cytomegalovirus promoter, with or without pseudotyping with VSV-G. VSV-G was placed under the control of polyhedrin promoter that is recognized by insect cells, allowing the formation of pseudotyped baculovirus. The study findings demonstrate that the pseudotyping of baculovirus significantly enhanced transduction efficiency compared to non-pseudotyped baculovirus, resulting in consequent distinction in the expression of GFP in mammalian cells. The results confirmed that pseudotyping is important for baculovirus mediated gene delivery. Further, when full-length VSV-G pseudotyping was compared with truncated VSV-G containing GED domain (G-stem of ectodomain in conjunction with the TM and CT domains of the glycoprotein), latter was relatively less efficient in transducing mammalian cells. This study demonstrated that pseudotyping with full-length VSV-G had better transduction efficiency in mammalian cells. However, at higher multiplicity of infection, both full-length and truncated VSV-G showed equivalent transduction. This study established the significance of pseudotyping of baculovirus with full-length VSV-G for efficient transduction of mammalian cells, utilizing the highly sensitive GFP marker system. These findings have significant implications in designing of baculovirus vector based antigen delivery for developing new generation vaccines.
摘要译文
重组杆状病毒构建了调查的VSV-G假型的必要性哺乳动物细胞转导这些病毒被设计巨细胞病毒启动的控制下表达绿色荧光蛋白（GFP）基因，有或没有用VSV-G假型VSV-G置于多角体蛋白启动子的控制是受昆虫细胞识别下，允许假型杆状病毒的形成研究结果表明，杆状病毒显著提高转导效率相比，非假型杆状病毒的假型，导致随之而来的区别在绿色荧光蛋白在哺乳动物cellsThe结果表达证实，假型是杆状病毒介导的基因deliveryFurther重要，当全长VSV-G假型与含有截短的VSV-G的GED域（G-茎胞外域的结合对TM和糖蛋白的CT结构域）相比，后者是相对的哺乳动物转导研究cellsThis效率较低表明，与全长VSV-G假型曾在哺乳动物cellsHowever更好的转染效率，在较高的感染复数，既全长和截短的VSV-G显示相当的转导本研究建立杆状病毒全长VSV-G的哺乳动物细胞高效转导的假的意义，利用高度敏感的GFP标记系统这些研究结果都在杆状病毒载体基于抗原传递设计开发的新一代疫苗显著影响
Sujit M. Kolangath [1] S. H. Basagoudanavar [1] M. Hosamani [1] P. Saravanan [1] R. P. Tamil Selvan [1]. Baculovirus mediated transduction: analysis of vesicular stomatitis virus glycoprotein pseudotyping[J]. VirusDisease, 2014,25(4): 441-446