【摘要】 目的了解扬州地区HPV16E7基因结构特点。方法从感染HPV16的宫颈癌组织中提取DNA,用PCR技术获得HPV16E7基因,以pGEM-Teasy为克隆载体构建重组质粒pGEM-T-E7,进行限制性核酸内切酶及核苷酸序列分析。结果扬州地方株HPV16E7基因与已报道的标准株(德国)仅在核苷酸序列上有一处突变,即199位的T→C,密码子由TTG变为CTG,而氨基酸序列未发生改变;与其他国家报道的HPV16E7核苷酸序列也存在一定差异,但同源性均在98.7%以上。结论扬州地方株HPV16E7基因的成功克隆,将丰富我国HPV16感染的流行病学资料,为HPV疫苗研制奠定基础。更多还原

【Abstract】 Objective: To investigate the gene sequence and mutations of HPV16 E7 in patients with HPV16 infection in the city of Yangzhou. Methods: Sample DNA was extracted from lesions in patients with cervical carcinoma resulted from HPV16 infection. The HPV16 E7 gene was amplified by PCR and inserted into the pGEM-Teasy vector to construct a recombinant plasmid pGEM-T-E7. Results: The sequence analysis showed that there was only one nucleotide mutation (T-C) in HPV16 E7, compared to the standard strain(German). The further amino acid analysis demonstrated that the mutation was from TTG to CTG. The homology of Yangzhou isolates with other strains of HPV16 E7 was above 98.7%. ~Conclusion : The successful cloning of the HPV16 E7 gene will enrich the epidemiologic data about HPV16 in China and make the foundation for HPV vaccine research.更多还原