%0 Journal Article
%T Study on the expression of Norovirus (GenegroupII) VP2 protein in insect cells and its subcellular localization<br>GII型诺如病毒VP2蛋白在昆虫细胞中的表达与定位研究
%A YAO Lin
%A KOU Xiao-Xi
%A JIANG Yan-Hu
%A WANG Lian-Zhu
%A WU Qing-Ping
%A ZHAI Yu-Xiu
%A <br>姚琳
%A 寇晓霞
%A 江艳华
%A 王联珠
%A 吴清平
%A 翟毓秀
%J 微生物学通报
%D 2012
%I 
%X Objective] This study is to express Norovirus (GenegroupII) VP2 in insect cells, analysis the subcellular localization of VP2 and lay the foundation of the function study of VP2. Methods] Norovirus (GenegroupII) ORF3 gene was amplified from plasmid pMD-ORF3 using primers P1/P2 (including 6×His tag coding sequence). The product was cloned into the vector pFastBac1 to construct recombinant plasmid pFB-ORF3. pFB-ORF3 was transformed into competent DH10Bac to get recombinant bacmid Bac-ORF3. Recombinant baculovirus, Ac-VP2, was generated for expressing VP2, by transfecting recombinant Bac-ORF3 with LipofactamineTM 2000 into sf9 insect cells. The expression product was testified by the western blot and indirect immunofluorescence assay with monoclonal antibody against 6×His tag. Results] The result of western blot showed a 29 kD specific bind in sf9 cells infected by Ac-VP2. Indirect immunofluorescence assay showed the specific green fluorescence in sf9 cells infected by Ac-VP2, and the green fluorescence could localize at nuclear and membrane of sf9 cells. Conclusion] These results demonstrated that the Norovirus (Gengroup II) VP2 was expressed in sf9 cells successfully and could localize at nuclear and membrane of sf9 cells.
%K Norovirus
%K VP2 protein
%K Insect cell
%K Expression
%K Subcellular localization<br>诺如病毒
%K VP2蛋白
%K 昆虫细胞
%K 表达
%K 亚细胞定位
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=78024727B5F4EF6AA9CA8E605B5FC464&aid=5EA8FDCC79E1D467607A8728CAEAE5C7&yid=99E9153A83D4CB11&vid=7C3A4C1EE6A45749&iid=38B194292C032A66&sid=DD54F1A968F26A72&eid=0E6E3FC0F9E9313C&journal_id=0253-2654&journal_name=微生物学通报&referenced_num=0&reference_num=0