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目的了解萍乡地方株HPV16 L1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中提取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。结果测定的序列与标准基因序列(NC Sbjct 496)比较,萍乡地区宫颈癌感染的HPV16型L1基因同源性为98.9%~99.6%,分别形成5种突变模式,其中3处由于核苷酸的改变,翻译的氨基酸也相应发生变化,但有1处由于核苷酸的丢失造成翻译的氨基酸也相应发生变化。结论萍乡地方株HPV16 L1核酸有一定程度的变异,可能会造成HPV16 L1蛋白免疫原性的差异。.此株改变也可能是本地区导致宫颈癌的主要原因。实验结果为研制适应本地区的基于HPV16 L1的基因工程疫苗提供了可靠实验依据。
Objective To understand the structural characteristics of HPV16 L1 gene in Pingxiang local strain and to analyze its polymorphism and protein characteristics of L1 gene. It provides a theoretical basis for the design and application of 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 fragments were ligated with T-vector and then introduced into E. coli DH-5α for plasmid DNA extraction Sequencing. Results Compared with the standard sequence (NC Sbjct 496), the homology of HPV16 L1 gene in cervical cancer in Pingxiang was 98.9% ~ 99.6%, which led to the formation of five mutation patterns, of which three were due to nucleotide Changes, translation of the amino acid changes accordingly, but there is a due to the loss of nucleotide translated amino acid also changed accordingly. Conclusion The HPV16 L1 nucleic acid in Pingxiang local strain has a certain degree of variation, which may result in the difference of immunogenicity of HPV16 L1 protein. This strain change may also be the main cause of cervical cancer in the region. The experimental results provide a reliable experimental basis for the development of HPV16 L1-based genetic engineering vaccine adapted to the region.