人乳头瘤病毒11型E7与共刺激分子CD80嵌合DNA疫苗质粒的构建及鉴定

Construction and identified of chimerical DNA vaccine plasmid of human papillomavirus type 11 E7 and CD80 co-stimulus gene

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 chimerical DNA vaccine plasmid of human papillomavirus type 11 （HPVⅡ） E7 and CD80 co - stimulatory gene. Methods： DNA was extracted from biopsy of condyloma tissues and subjected to PCR as template. The gene fragment of HPV Ⅱ -E7 was amplified by polymerase chain reaction （PCR） and was cloned into the pcDNA3. 1 （ ＋ ）. The recombinant plasmid pcDNA3. 1 （ ＋ ） / HPVⅡE7 was identified by restriction end nuclease cleanses digestion and sequencing. The gene fragment of HPVⅡ - E7 without terminator was amplified by PCR and was cloned into the pcDNA3. 1 （ ＋ ） /CD80 at the upstream of CD80. Results ： The amplified fragment from diseased tissue contained HPV Ⅱ- E7 gene was inserted into the vector of pcDNA3. 1 （ ＋ ） /CD80. Sequence in the constructed plasmids was correct. Conclusion： The chimerical DNA vaccine plasmid of pcDNA3. 1 （ ＋ ） /HPVⅡ E7 - CDS0 is constructed successfully, which may be used for further research of the fusion protein vaccine for the preventive and treatment of genital wart.