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通过PCR方法从人乳头瘤病毒 5 8型 (HPV5 8)全基因克隆中扩增出HPV5 8E6基因片段 ,克隆至真核表达质粒pcDNA3的T7启动子下游 ,并在体外转录成mRNA后 ,在源自网织红细胞的翻译缓冲液中成功表达了含有生物素标记的HPV5 8E6蛋白 ,并在体外成功发现HPV5 8E6能够降解p5 3蛋白 ,从而推断结合并降解p5 3蛋白是其致癌作用的关键环节 ,这为日后在细胞内对其致癌机理行进一步研究以及针对其进行治疗奠定下坚实的基础
The HPV5 8E6 gene fragment was amplified by PCR from the whole human papillomavirus type 5 (HPV58) gene and cloned into the downstream of the T7 promoter of the eukaryotic expression plasmid pcDNA3. After transcribed into mRNA in vitro, The successful expression of the biotinylated HPV5 8E6 protein from the reticulocyte translation buffer and the successful in vitro discovery that HPV5 8E6 degrades the p5 3 protein led to the inference that the combination and degradation of the p5 3 protein is a key part of its carcinogenicity, This provides a solid foundation for future research into its oncogenic mechanism in cells and its treatment