HSV-1 UL18基因重组载体的构建及其编码蛋白VP23的表达

Construction of recombinant vector of UL18 gene of HSV-1 and expression of its encoding protein VP23

目的:构建含有单纯疱疹病毒1型(HSV-1)UL18基因的原核表达载体。方法:先将UL18基因进行全序列PCR扩增,再将PCR产物克隆进pET-28a(+)质粒,最后将重组质粒转化进入原核表达系统E.coli BL21(DE3)中表达出带有6个组氨酸标签的重组VP23蛋白。结果:UL18基因正确重组进pET-28a(+)质粒,并在E.coli BL21(DE3)中表达。结论:成功构建单纯疱疹病毒1型(HSV-1)UL18基因的重组载体,并在原核表达系统表达出其编码的衣壳蛋白VP23且带有组氨酸标签,为进一步优化VP23在发酵中的最佳表达条件,纯化并大量制备VP23,以及制备VP23的多克隆抗体奠定了基础。

Objective： To construct the prokaryotic expression vectors containing UL18 gene. Methods： UL18 gene was amplified by PCR and cloned into plasmid pET - 28a（ ＋ ）. Then the recombinant plasmids were trans- formed into prokaryotic expressing system E. coli BL21 （ DE3 ） to express recombinant VP23 attaching with a 5 - His tag. Results： UL18 gene was correctly recombined into plasmid pET -28a（ ＋ ） , and expresses its encoding protein VP23 in E. coli BL21 （DE3）. Conclusion： The recombinant vectors containing UL18 gene were successfully constructed, and its encoding capsid protein VP23 attaching with a 6 - His tag was also expressed in prokaryotic expressing system, lying a foundation for further optimization of fermentation conditions, purification and preparation of VP23 polyclonal antibody.