HPV6晚期表达基因L1的克隆并在杆状病毒表达系统的表达及其抗血清制备

目的 获得具有正确序列的人乳头瘤病毒HPV6型晚期表达基因L1，并获得其高活性表达L1基因特异性抗血清，为进一步研究及临床检验应用奠定物质基础。方法 用PCR法从感染人乳头瘤病毒患者尖锐湿疣组织中获得HPV6晚期表达基因L1，测序验证后，将正确的HPV6晚期表达基因L1序列，插入Bac-to-Bac杆状病毒表达系统，转洒昆虫细胞Sf9，表达外壳蛋白L1，并以之作为抗原免疫兔，获得特异性抗血清。结果 获得了正确序列HPV6晚期表达基因L1，在Bac-to-Bac杆状病毒系统活性表达并获得其特异性抗血清。结论 获得HPV6晚期表达基因L1，及有抗原活性的表达和相应抗血清。

Cloning,Expression of HPV6 L1 in Bac-to-Bac Baculovlovirus Expression System and Preparation of anti-r-HPV6 L1 Serum

Objective Human Papillomavirus 6 L1 gene was cloned from condyloma acuminata and expressed actively in Sf9 insect cells by Bac-to-Bac Baculovirus expression vector system.Methods Human Papillomavirus6 L1 gene was acquired by PCR from CA.At first,Human Papillomavirus type6 L1 gene was insert into pFast Bac vector,then the recombiant pFast Bac-Angio(K1-4) transfect competent DH10 Bac cells to construct recombiant HPV6 L1 Bacmid DNA,Sf9 cells were cotrasfected with recombiant HPV6 L1-Bacmid DNA using cellfectin.Then recombiant HPV6 L1 was used as antigen to immunity rabbit.Results After 4 round amplification of recombiant baculovirus,high expression level of HPV6 L1 in Sf9 cells infected was detected by immunoblotting with HPV6 L1 antibody,and anti-reHPV6 L1 was acquired.The usitility in diagniosis and therapy need further study.Conclution HPV6 L1 genewas acquired and actively expressed in Bac-to-Bac Baculovirus expression system.The anti-HPV6 L1 serum has been acquired.