中国人兽共患病学报2011，Vol.27Issue(5)：376-380,5.
登革2型病毒包膜蛋白在毕赤酵母中的表达和病毒样颗粒的组装
Expression and self-assembly of virus-likes particles containing dengue virus type 2 envelope proteins in Pichia pastoris
摘要
Abstract
The aim of this study was to express the dengue type 2 envelope and pre-membrane proteins in the constitutive Pichia pastoris system, and study the assembly characteristic of virus-like particles in cells. The virus mRNA was taken as the template to amplify the prM/E genes. Then the genes were cloned into the multiple clone sites of the expression vector pGAPZa A,and transformed into Pichia pastoris GS115. By the selection of Zeocin and the identification of PCR, the positive clones was expressed and analyzed. Results demonstrated that dengue virus type 2 prM/E genes were successfully cloned into expression vector; SDS-PAGE and Western blot showed that the envelope protein was high-level expressed in constitutive Pichia pastoris system; the virus-like particles with diameters of 30-50 nm, and the double membrane structures which was similar with the cells infected dengue virus were detected by transmission electron microscopy; results of immune-electron microscopy indicated that the recombinant virus-like particles could be recognized by the anti-dengue type 2 mouse serum, revealed that the particles had immunoreactivity. These results indicate that the constitutive Pichia pastoris system which express dengue type 2 virus-like particles has been established successfully,which will provide the basis for further studies of dengue virus-like particles vaccine.关键词
登革病毒/包膜蛋白/毕赤酵母/病毒样颗粒Key words
dengue virus/ envelope protein/ Pichia pastoris/ virus-like particles分类
医药卫生引用本文复制引用
刘文权,江丽芳,刘岩,江汉宁,汤云霞,方丹云..登革2型病毒包膜蛋白在毕赤酵母中的表达和病毒样颗粒的组装[J].中国人兽共患病学报,2011,27(5):376-380,5.基金项目
国家自然科学广东省联合基金(U0632002)和国家高技术研究发展计划"863"计划项目(2006AA02A223)联合资助 （U0632002）
