EB病毒包膜糖蛋白gp85真核表达载体的构建  

Construction of Expression Vector for Membrane Glycoprotein gp85 of EB Virus

作　　者：连云宗[1] 李极品[1] 吴玉水[1] 程烽[1] 朱忠勇[1] 

机构地区：[1]南京军区福州总医院全军医学检验中心,福建福州350025

出　　处：《生物技术通讯》2005年第3期276-277,共2页Letters in Biotechnology

基　　金：福建省青年科技人才创新项目(2003J056)

摘　　要：用基因工程技术克隆EB病毒中抗原性较强的膜蛋白gp85的编码基因BXLF2,构建真核表达载体。以EB病毒B95-8细胞培养上清为模板,PCR扩增出BXLF2基因。PCR产物经SnaBⅠ和NotⅠ双酶切后克隆至毕赤酵母表达载体pPIC9K,用双酶切和DNA测序鉴定重组质粒。重组质粒双酶切的片段大小与预期符合,重组克隆外源基因的测序结果与文献报道一致。结果表明,EB病毒gp85的编码基因BXLF2被成功地克隆入真核表达载体pPIC9K,为下一步在毕赤酵母中表达EB病毒gp85蛋白建立了基础。The BXLF2 gene was PCR-amplified from culture supernant of B95-8 cell line. The PCR product was digested with SnaBⅠand NotⅠ,and cloned into expression vector pPIC9k for the yeast Pichia pastoris. The constructed recombinant was confirmed by restriction enzyme analysis and DNA sequencing. The BXLF2 gene was successfully amplified using PCR. Double digestion of the constructed plasmid revealed DNA fragments of expected size. The sequence of the inserted BXLF2 gene was identical to that of published. The BXLF2 gene coding gp85 of EB virus was cloned into pPIC9K, which should provide the condition for expressing the protein in P. pastoris.

关 键 词：真核表达载体 EB病毒 包膜糖蛋白 构建 基因工程技术 酵母表达载体 编码基因 重组质粒 PCR扩增 PCR产物 DNA测序 双酶切 细胞培养 文献报道 外源基因 重组克隆 毕赤酵母 膜蛋白 抗原性 

分 类 号：Q782[生物学—分子生物学] Q939.4