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目的 构建重组原核表达质粒以获得HPV16L1活性蛋白 ,为进一步研制HPV16基因工程疫苗打下基础。方法 以克隆质粒pCR2 .1-HPV16L1为模板 ,用PCR方法扩增HPV16L1DNA片段 ,利用pQE31质粒载体构建pQE31 HPV16L1重组质粒 ,用酶切电泳验证重组结果的正确性 ,测序检查质粒重组后序列情况。结果 PCR结果显示扩增片断大小约 1.5Kb ,与预期相同。重组质粒酶切后显示其大小约 5 .0Kb。大小及酶切图谱与预期相同。经测序发现插入片段两端序列无改变。结论 pQE31 HPV16L1质粒构建成功。
Objective To construct recombinant prokaryotic expression plasmid to obtain HPV16 L1 active protein, which laid the foundation for further development of HPV16 gene engineering vaccine. Methods The cloned plasmid pCR2 .1-HPV16L1 was used as a template to amplify the HPV16L1 DNA fragment by PCR. The pQE31 HPV16L1 recombinant plasmid was constructed by pQE31 plasmid vector. The correctness of the recombinant plasmid was confirmed by restriction endonuclease digestion and sequencing. Results PCR results showed that the amplified fragment size of about 1.5Kb, the same as expected. Recombinant plasmid showed its size after digestion of about 5 .0Kb. Size and restriction map with the same expectations. Sequencing showed no change in the sequence of the inserted fragment. Conclusion pQE31 HPV16L1 plasmid was successfully constructed.