人乳头瘤病毒6型E7基因完整开放阅读框架的克隆和鉴定  

Cloning and Identification of Complete Open Reading Frame of E7 Gene of Human Papillomavirus Type 6

作　　者：丛宪玲[1] 田亚萍[2] 陈越[1] 王丽颖[2] 于永利[1] 

机构地区：[1]吉林大学基础医学院免疫学教研室 [2]吉林大学基础医学院分子生物学教研室

出　　处：《中国实验诊断学》2005年第6期868-870,共3页Chinese Journal of Laboratory Diagnosis

基　　金：国家重大基础科研基金973项目资助(001CB510007)

摘　　要：目的从尖锐湿疣病变组织中克隆人乳头瘤病毒6型(HPV6)E7基因,为HPV感染的检测及基因工程疫苗的研制奠定基础。方法以临床尖锐湿疣标本总DNA为模板,采用聚合酶链反应(PCR)技术,从尖锐湿疣病变组织中扩增出人乳头瘤病毒6型E7基因完整开放阅读框架,并插入载体PMD-18T中构建重组质粒,转化JM109中扩增,进行酶切及测序分析。结果测序结果证实克隆成功。结论该实验为研究人乳头瘤病毒E7蛋白的免疫学和流行病学特点创造了条件,也为下一步制备HPV6型疫苗奠定了基础。Objective To construct DNA vaccine against human papillomavirus type 6. target gene E7 was cloned from biopsy of condyloma tissues. Methods HPV DNA was subjected to PCR as template. The PCR product was inserted into PMD-18T vector and the recombinant plasmid PMD-18T-HPV6E7 was identified by endonuclease restriction digestion and sequencing. Results A BLAST search confirmed that the cloned fragment contained the complete open reading frame of E7 gene of HPV 6. Conclusion The target gene may provide a ready source for HPV vaccine preparation.

关 键 词：人乳头瘤病毒 基因克隆 聚合酶链反应 序列分析 

分 类 号：R373[医药卫生—病原生物学]