尖锐湿疣组织HPV6b及11型L1基因多态性及蛋白特性分析.doc

尖锐湿疣组织HPV6b及11型L1基因多态性及蛋白特性分析 　 作者：欧琴，石朝辉，朱珊丽，张丽芳 【摘要】 目的:调查温州地区尖锐湿疣组织HPV6b和11型感染情况，分析其L1基因多态性及蛋白特性变化，为基因工程重组L1蛋白疫苗的应用提供理论依据。方法:采用HPV6b和11型L1基因特异性引物对31份温州地区尖锐湿疣组织标本进行PCR扩增，并各取7份阳性PCR产物直接测序，用软件分析HPV6b和11型L1基因多态性及蛋白特性。结果:31份标本中，HPV6b和11型阳性率分别为35.5%和45.2%。与标准基因序列比较，HPV6b和11型L1基因分别形成5和6种突变模式，同源性分别为99.8%～99.9%和99.7%～99.9%。HPV6b型L1基因中仅1处碱基突变引起所编码氨基酸的变异(S→P)，但其编码蛋白在亲水性、表面可及性及抗原性上无明显变化；HPV11型L1基因的碱基突变均为无义突变，所编码氨基酸与标准株完全一致。结论:HPV6b和11型是温州地区尖锐湿疣的主要感染型别；HPV6b和11型L1基因序列高度保守。 【关键词】 人乳头瘤病毒；尖锐湿疣；L1基因；突变；基因多态性 Abstract: Objective: To study the viral types of Human Papillomavirus (HPV) in condylomata acuminate from Wenzhou area，and to analyze the changes of L1 gene polymorphism and protein property.Methods: The tissue DNA was extracted from 31 biopsy specimens of condylomata acuminate. The L1 gene of HPV was amplified with PCR by 6b and 11 type specific primers respectively. And seven positive L1 PCR products of each type were identified by sequencing.At last， it was L1 gene polymorphism and protein property that were analysied by biosoft. Results: The results of PCR showed that the positive rate of HPV6b and 11 L1 were 35.5 % and 45.2% respectively. Compared with standerd L1 gene sequence， five mutations were found and gene homology were 99.8%～99.9% in HPV6b type，six mutations were found and gene homology were 99.7%～99.9% in HPV11 type.The gene mutation of HPV6b L1 from S to P formed the change at nucleic acid level，but there was no significant difference in hydrophilicity，surface probality and antigencity. And the gene mutations of HPV11 L1 were all nonsense. Conclusion: HPV6b and 11 are the main infected types in the Wenzhou area， and the L1 gene sequence of HPV6b and 11 is highly conservative. Key words: human papillomavirus；condylomata acuminate；L1 genes；mutation；gene polymorphism 尖锐湿疣(condylomata acuminata，CA)主要由人乳头瘤病毒( human papillomavirus，HPV) 6型和11型经性传播感染引起[1]。HPV L1基因可在杆状病毒-昆虫细胞系统中自行组装成病毒样颗粒(VLPs)，这种不含病毒DNA的空衣壳具有较强的抗原性和免疫原性，可刺激B细胞产生中和抗体，阻断病毒感染，因而L1蛋白成为了基因工程疫苗研究的重要靶抗原。