HPV58型衣壳蛋白L1和L2基因的克隆表达及病毒样颗粒的装配

目的大量表达和纯化HPV58衣壳蛋白,形成病毒相似颗粒。方法制备携带HPV58L1和L2基因的重组杆状病毒,转染昆虫细胞,在真核细胞中大量表达HPV58L1和L2蛋白并进行纯化。电镜观察病毒相似颗粒的形成情况。免疫小鼠制备特异性抗血清。结果 HPV58L1和L2蛋白在真核细胞中被高效表达,电镜下见表达纯化的病毒衣壳蛋白L1与L2共同组装形成病毒相似颗粒(VLPs),用其免疫小鼠能产生针对HPV58L1蛋白的高滴度的特异性抗体。结论在真核细胞中大量表达HPV58L1和L2蛋白,纯化后能形成病毒相似颗粒,并具备很强的免疫原性,有望用其制作宫颈癌预防性疫苗。

Gene Cloning and Expression of HPV58 Capsid Protein L1 and L2 and the Virus-Like Particles Assembling

Objective Express and purificate HPV58 capsid protein quantity,to make it to virus-like particles.Mothods Prepare recombination Baculovirus which phore HPV58L1 and HPV58L2 genic,transfection entomo-cyte,express HPV58L1 and HPV58L2 protein in the ucaryotic cell quantity and purificate simultaneous.Use the ectron microscope to observate the condition of the formation of the virus-like particles.Immunity mouse prepare the specific antisera.Results HPV58L1 and HPV58L2 protein are high expressed in the eucaryotic cell,by the ectron microscope,we can see the purification viral capsid protein L1 and L2 are concomitance compose the virus-like particles（VLPs）,use its immunity mouse can productive the high titer specific antibody what to aim directly at HPV58L1 protein.Conclusion HPV58L1 and HPV58L2 protein can be high expressed in the ucaryotic cell,after purificated they can productive the virus-like particles,meanwhile,they are provided with powerful immunogenicity.They are promisingly to be maked the prophylactic vaccine of the uterine cervix cancer.