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目的了解萍乡地区地方株HPV16L1基因结构特点,分析其结构蛋白L1基因多态性及蛋白特性变化,为基于L1基因工程疫苗的设计和应用提供理论依据。方法设计HPV16型L1基因特异性引物,从宫颈脱落上皮细胞提取DNA,进行PCR扩增鉴定,将扩增的片段同T-载体连接,然后导入到大肠杆菌DH-5α中培养,提取质粒DNA进行测序。结果将测序结果导出数据后,根绝NCBI网站上公布的GENBANK数据库,利用BLAST工具将9个样本的序列同相应的标准株序列进行比对,9个HPV16样本同标准株的同源性分别为98.9%～99.6%。结论 9株萍乡地方株HPV16L1基因与标准株之间的差异,特别可能会造成HPV16L1蛋白免疫原性的差异。实验结果可为研制适应本地区的基于HPV16L1的基因工程疫苗提供可靠实验依据。
Objective To understand the structural characteristics of HPV16 L1 gene in the local strain of Pingxiang and to analyze the polymorphism of its structural protein L1 gene and the change of its protein characteristics, so as to provide a theoretical basis for the design and application of the L1 gene engineering vaccine. Methods HPV16 type L1 gene-specific primers were designed. DNA was extracted from cervical exfoliated epithelial cells and identified by PCR. The amplified fragment was ligated with T-vector and then introduced into E. coli DH-5α for cultivation. The plasmid DNA was extracted Sequencing. Results After the data were exported from the sequencing results, the GENBANK database published on the NCBI website was eradicated. The sequences of 9 samples were aligned with the corresponding standard strains using the BLAST tool. The homologies of the 9 HPV16 samples to the standard strains were 98.9 % ~ 99.6%. Conclusion The differences between the HPV16 L1 gene of 9 Pingxiang local strains and the standard strains may cause the difference of the immunogenicity of HPV16 L1 protein in particular. The experimental results can provide a reliable experimental basis for the development of HPV16L1-based genetic engineering vaccine adapted to the region.