【摘要】 人乳头瘤病毒(HPV)16型感染与宫颈癌的发生、发展密切相关,故研制HPW16型基因工程疫苗对预防HPV感染、控制宫颈癌的发生有重要意义。本研究构建HPV16 L1基因工程菌并进行了基础实验研究。首先采用PCR技术从3例宫颈癌组织中扩增和克隆了HPV16L1基因片段并进行测序。进而构建了pPIC3.5～HPV16L1真核表达工程菌、pQE31～HPV16L1和pGEX4T-HPV16LI原核表达工程菌及杆状病毒-昆虫细胞表达系统。经SDS-PAGE、Westem blot和电镜等手段证明了工程菌表达HPV16 L1蛋白的分子特性和免疫原性,而且,重组菌表达的HPV16 L1可以形成类病毒颗粒(VLP)。经免疫BALB/C小鼠可获得高效价特异血清,证实重组HPV16 L1蛋白具有良好的免疫原性。为进一步研制临床应用疫苗及血清学诊断试剂提供了实验基础。更多还原

【Abstract】 Human papillomavirus (HPV) type 16 is highly related to the initiation and development of human cervical cancer, therefore, it is very important to developing recombinant vaccine of HPV16 for the prevention and control of HPV 16 infection and cervical cancer.The re-combinant vaccines based on HPV16 L1 gene were founded and experimental studied in this project. Firstly, HPV16 L1 gene was amplified, cloned and sequenced from three cervical cancer tissues by polymerase chain reaction (PCR) .Eukaryoticc expression system pPIC3.5-HPV16 L1, prokaryotic expression systems pQE3.1 - HPV16 L1 and pGEX4T - HPV16 L1, and a baculiform virus - insect cell expression system were constructed.The molecular specificities and immunogenicity of those recombinant L1 proteins were confirmed by means of SDS - PAGE, western blot and electromicroscopy.The recombinant L1 protein could form virus-like particle (VIP) . High - titer antibody were obtained by inoculating BALB/C mouse with the recombinant L1 proteins, it showed that the recombinant L1 proteins preserved excellent immunogenicity.Thisproject provides possibility for further work on clinic applicable HPV16 vaccine and diagnostic reagents.更多还原