pQE31-HPV16 L1重组质粒的构建及鉴定

Construction and identification of recombinant plasmid pQE31-HPV16 L1

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 a recombinant plasmid pQE31 HPV16 L1 for obtaining the HPV16 L1 protein expressed in E.coli and developing the genetically engineered vaccine.Methods PCR was used to amplify the HPV16 L1 gene from plasmid pCR2.1 HPV16 L1 in which the HPV16 L1 was cloned. A new plasmid, pQE31 HPV16 L1, was then constructed by inserting the amplified HPV16 L1 gene into pQE31, a prokaryotic expression vector. Restriction analysis and sequencing were used to confirm the structure of pQE31 HPV16 L1.Results A DNA fragment in size of 1.5Kb was amplified. Restriction analysis showed that the amplified gene was inserted into pQE31 correctly. Sequencing showed there was no mutation in both ends of the inserted L1 gene.Conclusion The plasmid pQE31 HPV16 L1 is constructed successfully.