张飞齐;杨露;杨晓明;

• 1:武汉生物制品研究所
• 2:中国生物技术股份有限责任公司

摘要(Abstract)：

目的建立人乳头瘤病毒(human papillomavirus,HPV)16型嵌合病毒样颗粒(virus-like particle,VLP)体外形成实验的方法。方法以HPV16型L1为模板,将两个L2保守序列(aa56⁷5,aa10875,aa108¹20)分别插入L1两个可插入位点(DE袢状区,h4状区),构建出4种嵌合蛋白基因,利用Bac-to-Bac杆状病毒-昆虫表达系统表达嵌合的4种VLP蛋白,经蔗糖密度梯度离心纯化,纯化的目的蛋白进行VLP体外解聚重聚实验,Western blot分析体外解聚重聚前后的蛋白。结果表达及纯化的4种HPV16 L1-L2嵌合蛋白相对分子质量均约60 000,大小与预期相符;经VLP体外解聚重聚实验,使不能很好自组装的4种HPV16 L1-L2嵌合蛋白组装形成VLP,并具有免疫原性。结论建立的方法有助于HPV嵌合VLP的形成,能够明显提高其组装效率,为以L2蛋白为目标的第二代HPV疫苗的嵌合VLP体外组装提供了可行的实验方法。

关键词(KeyWords)： 人乳头瘤病毒16型;基因重组;嵌合病毒样颗粒;体外重组实验

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 张飞齐;杨露;杨晓明;

DOI: 10.13200/j.cnki.cjb.002286

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