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目的:构建人乳头瘤病毒11型(HPV ll)E7与共刺激分子CDS0嵌合DNA疫苗质粒。方法:从尖锐湿疣病变组织中提取DNA制备模板,采用聚合酶链反应(PCR)技术扩增HPV ll-E7基因,经双酶切后定向克隆到pcDNA3.1(+)中,获得pcDNA3.1(+)/HPV llE7,测序鉴定后,再用PCR方法扩增去除终止密码子的E7基因片段,按上述方法插入质粒pcD-NA3.1(+)/CD80中CD80上游,构建pcDNA3.1(+)/HPV llE7-CD80融合基因真核表达质粒。结果:去除终止子的E7基因片段成功正向插入CD80上游,双向测序分析显示序列无误,pcDNA3.1(+)/HPV llE7-CD80嵌合真核表达质粒构建成功。结论:HPV ll-E7/CD80嵌合DNA疫苗质粒构建成功,为研究该疫苗在尖锐湿疣防治中的作用奠定了基础。
Objective: To construct human papillomavirus type 11 (HPV11) E7 and costimulatory molecule CDS0 chimeric DNA vaccine plasmid. Methods: The genomic DNA was extracted from the condyloma acuminatum tissues. The HPV11-E7 gene was amplified by polymerase chain reaction (PCR) and cloned into pcDNA3.1 (+) vector to obtain pcDNA3.1 (+) / HPV llE7. After sequencing, the E7 gene fragment with the stop codon was amplified by PCR and inserted into the upstream of CD80 plasmid pcD-NA3.1 (+) / CD80 as above to construct pcDNA3.1 +) / HPV llE7-CD80 fusion gene eukaryotic expression plasmid. Results: The E7 gene fragment without terminator was successfully inserted into the upstream of CD80 successfully. The two-way sequencing showed that the sequence was correct. The construction of pcDNA3.1 (+) / HPV11E7-CD80 chimeric eukaryotic expression plasmid was successful. CONCLUSION: The construction of HPV ll-E7 / CD80 chimeric DNA vaccine plasmid has been successfully constructed, which lays the foundation for the study of the vaccine in the prevention and treatment of condyloma acuminatum.