洪敏, 曾丽雯, 梁燕, 杨子江, 李琦涵. Ⅰ型单纯疱疹病毒包膜糖蛋白L基因片段的克隆及高效表达[J]. 云南大学学报(自然科学版), 2007, 29(2): 194-198.
引用本文: 洪敏, 曾丽雯, 梁燕, 杨子江, 李琦涵. Ⅰ型单纯疱疹病毒包膜糖蛋白L基因片段的克隆及高效表达[J]. 云南大学学报(自然科学版), 2007, 29(2): 194-198.
HONG Min, ZENG Li-wen, LIANG Yan, YANG Zi-jiang, LI Qi-han. Cloning and expression of herpes simplex virus type 1 glycoprotein L gene[J]. Journal of Yunnan University: Natural Sciences Edition, 2007, 29(2): 194-198.
Citation: HONG Min, ZENG Li-wen, LIANG Yan, YANG Zi-jiang, LI Qi-han. Cloning and expression of herpes simplex virus type 1 glycoprotein L gene[J]. Journal of Yunnan University: Natural Sciences Edition, 2007, 29(2): 194-198.

Ⅰ型单纯疱疹病毒包膜糖蛋白L基因片段的克隆及高效表达

Cloning and expression of herpes simplex virus type 1 glycoprotein L gene

  • 摘要: 采用PCR方法扩增HSV-1病毒型特异性包膜糖蛋白L(gL)基因片段并克隆至原核表达载体pGEX-5X-1获得重组质粒pGEX-5X-1-gL,将重组质粒转化E.coli BL21表达菌后经IPTG诱导表达目的蛋白.SDS-PAGE蛋白检测表明,在分子质量56 ku处有HSV-1 GST-gL融合蛋白的高效表达,通过IPTG浓度筛选和诱导前表达菌扩增培养时间的比较分析对诱导条件进行了优化,GST-gL融合蛋白表达量可达到菌体蛋白总量的48.65%.Western blot中利用HSV-1灭活病毒获得的多克隆抗体确证所表达蛋白为HSV-1病毒组分.这一表达系统的建立和优化对进一步探讨HSV-1 gL蛋白功能及其免疫原性提供了有利条件.

     

    Abstract: The herpes simplex virus type 1(HSV-1)glycoprotein L(gL)gene was amplified by PCR and cloned into the prokaryotic expressive vector pGEX-5X-1.The recombinant vector pGEX-5X-1-gL was identified by PCR,restriction digestion and sequence analysis.GST-gL fusion protein was expressed efficiently in E.coli BL21 though IPTG as an inducer.The fusion protein band with size of 56 ku was detected by SDS-PAGE.An optimal expression system was established though screen IPTG concentration and inducing procedure,which yielded GST-gL fusion protein.It is 48.65 percent of total protein expressed in E.coli BL21.The HSV-1 polyclonal antibodies were produced by using HSV-1 virus.Western blot assay shows these polyclonal antibodies certified the fusion protein as a component of HSV-1 virus.A good foundation to discuss gL protein biological function and immunogenicity in future are provided.

     

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