单纯疱疹病毒2型感染细胞多肽27真核表达质粒的构建及表达

Construction and Expression of Eukaryotic Expression Plasmid for ICP27 of Herpes Simplex Virus Type 2

为了构建单纯疱疹病毒2型(HSV-2)感染细胞多肽27(ICP27)真核表达质粒,应用PCR技术从HSV-2 333株的基因组中扩增ICP27基因,并连接至真核表达载体pEGFPC2,对阳性克隆进行菌落PCR、酶切和测序鉴定后,成功构建了重组质粒pEGFPC2-ICP27。用X fect转染试剂盒将重组质粒pEGFPC2-ICP27转染至Vero细胞中,并用RT-PCR及W estern b lot-ting检测其表达情况。结果显示,ICP27基因在Vero细胞中得到正确表达。真核表达质粒pEGFPC2-ICP27的构建成功,为进一步研究ICP27对宿主细胞的影响奠定了基础。

To construct the eukaryotic expression plasmid for infected-cell polypeptide 27（ICP27）and detect its expression in Vero cells,the sequence of ICP27 was cloned by PCR and then the PCR products were inserted into pEGFPC2 to construct a recombinant plasmid.The constructed recombinant plasmid was finally transfected into Vero cells with Xfect for expression after the right sequence was confirmed by restrictive endonuclease analysis and DNA sequencing.The expression of ICP27 was detected in the lysate of transfected Vero cells by RT-PCR and western blotting.The successful construction of eukaryotic expression plasmid pEGFPC2-ICP27 has laid the foundation of further studying on the effect of ICP27 on host cells.