基于抗原表位分析的人乳头瘤病毒16L1壳蛋白基因克隆及表达纯化

发布时间：2018-07-06 10:28

本文选题：表位分析 + HPVL　；参考：《现代妇产科进展》2015年07期

【摘要】：目的:选择人乳头瘤病毒(HPV)16L1抗原性最强的一段抗原表位编码区,构建原核表达载体并利用大肠杆菌表达抗原蛋白,为HPV疫苗的研究及宫颈癌的诊断奠定基础。方法:借助DNAstar软件分析HPV16L1的CDS区,选择抗原性最强的一段抗原表位编码区,提取HPV16L1编码区的DNA。目的 DNA和p ET32a质粒分别进行双酶切,连接并转化DH5α大肠杆菌,筛选阳性克隆后转化BL21大肠杆菌,诱导产生HPV16L1重组蛋白,通过市售抗体对重组蛋白的抗原特异性进行鉴定。结果:利用DNAstar软件,选择了抗原表位较富集的碱基序列919~1314bp区段,按常规制备重组蛋白的方法成功制备HPV16L1重组蛋白。抗体鉴定表明,制备的重组蛋白抗原特异性良好。结论:有针对性地制备HPV16L1重组蛋白,可避免其他抗原表位的干扰。经鉴定,HPV16L1重组蛋白抗原特异性良好,将为HPV疫苗的研究及宫颈癌的诊断奠定基础。
[Abstract]:Aim: to construct the prokaryotic expression vector of human papillomavirus (HPV) 16L1 antigen epitope coding region and to express antigen protein by E. coli, so as to lay a foundation for the study of HPV vaccine and the diagnosis of cervical cancer. Methods: the CDS region of HPV16 L1 was analyzed by DNAstar software, and the most antigenic epitope coding region was selected, and the DNAs of HPV16 L1 coding region were extracted. Objective DNA and pET32a plasmids were digested respectively, ligated and transformed into DH5 伪 Escherichia coli, then transformed into BL21 Escherichia coli after screening positive clones, and induced the production of HPV16L1 recombinant protein. The antigen specificity of the recombinant protein was identified by market antibody. Results: the recombinant protein HPV16 L1 was successfully prepared by using DNAstar software to select the 919~1314bp region of the base sequence enriched by antigen epitope and to prepare the recombinant protein according to the routine method. Antibody identification showed that the prepared recombinant protein antigen had good specificity. Conclusion: the preparation of HPV16 L1 recombinant protein can avoid the interference of other antigenic epitopes. The specificity of HPV16 L1 recombinant protein antigen is good, which will lay a foundation for the study of HPV vaccine and the diagnosis of cervical cancer.
【作者单位】：广州医科大学附属广东省妇女儿童医院妇科;广州医科大学附属广东省妇女儿童医院麻醉科;
【分类号】：R737.33

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