人乳头瘤病毒16型和11型L1基因双价重组杆状病毒转移质粒的构建及鉴定

目的：构建人乳头瘤病毒16型与11型L1基因双价重组杆状病毒转移质粒并对其进行鉴定。方法：采用：PCR法从尖锐湿疣组织标本中扩增人乳头瘤病毒11型晚期基因L1，并对其进行克隆测序；利用基因重组技术将人乳头瘤病毒16型和11型L1晚期基因共同装入杆状病毒转移载体中，分别位于强启动子Ppolh和弱启动子P10之下；利用酶切和PCR技术对双价重组杆状病毒转移质粒进行鉴定。结果：PCR扩增法获得尖税湿疣组织中感染的人乳头瘤病毒11型的L1基因，得到人乳头瘤病毒16型L1和11型L1基因双价重组杆状病毒转移质粒，经酶切电泳鉴定验证重组成功。结论：本研究成功构建了双价重组杆状病毒转移质粒，为进一步构建双价L1蛋白表达系统进而建立双价基因工程亚单位疫苗打下基础。

CONSTRUCTION AND IDENTIF ICATION OF HUMAN PAPILLOMAVIRUS TYPE 16 AND 11 L1 GENE BIVALENT RECOMBINANT BACULOVIRUS TRANSFER PLASMID

Objective:To construct and identify th e bivalent recombinant baculovirus transfer plasmid of HPV type 16 and type 11 gen e. Methods:HPV11 late L1 was amplified from Condyloma acuminate t i ssue by the polymerase chain reaction(PCR).The amplified products of the HPV11 L1 fragments were coloned and sequenced.The HPV16 and 11 L1 were inserted into th e baculovirus transfer vector pFastbac-Dual,16 L1 was followed powerful promoto r Ppolh and 11 L1 followed P10 weak promotor.The bivalent recombinant baculovirus transfer plasmid were characterized by enzyme digesting.Results:The HPV type 11 L1 gene in Condylomata acuminatum tissu e were obtained.The bivalent recombinant baculovirus transfer plasmid of HPV typ e 16 and type 11 L1 gene was constructed and characterized successfully.Conclusion:The bivalent recombinant baculovirus transfer plasmi d will be applied to the constuction of bivalent L1 protein expression system,th en contributed to the establishment the bivalent gene engineering subunit vaccin e.