猕猴B病毒gB蛋白胞外区基因片段的合成和真核表达  

Synthesis and eukaryotic expressing of the gene encoding extracellular domains of gB protein B virus

作　　者：刘慧芳[1,2] 孙书芳 曾林[1] 易思萌 游颖[1] 马雨楠 范君文[1] 孙兆增[1] 王新[2] 

机构地区：[1]军事医学科学院实验动物中心,北京100071 [2]青岛农业大学,山东青岛266109 [3]山东省青岛市中心血站,山东青岛266071

出　　处：《中国比较医学杂志》2014年第11期6-9,I0003,共5页Chinese Journal of Comparative Medicine

基　　金：国家科技支撑计划(2011BAI15B03);十二五重大传染病专项(2011ZX09201-031)

摘　　要：目的获得猕猴B病毒gB蛋白胞外区基因,并分析其表达特性。方法利用基因合成的方法合成B病毒gB蛋白胞外区的基因片段,经BamHⅠ和NotⅠ双酶切后连接到pEGFP-N3载体。将构建的pEGFP-N3-GB合重组质粒转染到293细胞。提取蛋白用Western blot检测其在细胞内的表达情况,并利用激光共聚焦分析在细胞内的表达定位。结果 pEGFP-N3-GB合重组质粒能在293细胞内正常表达,并且在细胞膜上表达。结论利用真核表达系统,可以产生B病毒gB蛋白的特异性抗原重组蛋白,且在细胞膜上表达。Objective To get the gene encoding extracellular domains of gB protein of B virus and analyze its expression in the eukaryocyte cell. Methods synthesizing gene fragment encoding extracellular domains of gB protein of B virus was by using synthesis gene, then digested with the restriction endonucleases BamH I and Not I and inserted into eukaryotic expressing vector pEGFP-N3, pEGFP-N3-GB合 was transfectcd into 293 cells. After protein extraction, the expression of gene was detcted by western blotting, and the cellular localization of the gene was analyzed by immunofluorescence and laser scanning confoeal microscopy. Results pEGFP-N3-GB合 were expressed in 293 ceils and on the cell membrane. Conclusion eukaryotic expressing system can produce specific antigen recombination protein of B virus gB protein and express on the cell membrane.

关 键 词：猕猴B病毒 gB蛋白 合成 真核表达 

分 类 号：R332[医药卫生—人体生理学]