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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。
Human papillomavirus (HPV) 16 infection is closely related to the occurrence and development of cervical cancer. Therefore, it is of great significance to develop HPW16 genetic engineering vaccine to prevent HPV infection and control the occurrence of cervical cancer. This study constructed HPV16 L1 gene engineering bacteria and carried out the basic experimental study. First, HPV16L1 gene fragment was amplified and cloned from 3 cervical cancer tissues by PCR and sequenced. Then we constructed the eukaryotic expression plasmid pPIC3.5 ~ HPV16L1, pQE31 ~ HPV16L1 and pGEX4T-HPV16LI prokaryotic expression baculovirus and baculovirus-insect cell expression system. The molecular characteristics and immunogenicity of HPV16 L1 protein were demonstrated by SDS-PAGE, Westem blot and electron microscopy. Moreover, HPV16 L1 expressed by recombinant bacteria could form virus-like particle (VLP). High titer specific serum was obtained from the BALB / C mice immunized and confirmed that the recombinant HPV16 L1 protein has good immunogenicity. It provided the experimental basis for further development of clinical application of vaccine and serological diagnostic reagents.