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目的分析中国山东地区宫颈癌患者ＨＰＶ１６型Ｅ６Ｅ７基因结构特点。方法从中国山东地区宫颈癌活检组织中提取ＤＮＡ，经ＨＰＶ多重引物ＰＣＲ法鉴定标本中感染ＨＰＶ型别，选感染ＨＰＶ１６型的标本ＤＮＡ为模板进行ＰＣＲ扩增，获得ＨＰＶ１６Ｅ６Ｅ７基因，将其重组入ｐＡＬＴＥＲ－１载体，进行双向测序、分析。结果构建了含中国山东地区宫颈癌患者组织中ＨＰＶ１６Ｅ６Ｅ７的重组质粒，命名为ＨＰＶ１６Ｅ６Ｅ７－ＳＤ。ＤＮＡ序列分析表明，该序列全长７７６ｂｐ，与已发表的德国标准株长度相等，但其核苷酸顺序的第５５７位核苷酸“Ｔ”变为“Ｃ”，即Ｅ６的终止密码子ＴＡＡ变为谷氨酰胺密码子ＣＡＡ。结论中国山东地区宫颈癌患者组织中ＨＰＶ１６Ｅ６Ｅ７的基因结构与德国标准株ＨＰＶ１６Ｅ６Ｅ７基因之间存在差异。
Objective To analyze the structural characteristics of HPV16 E6E7 gene in patients with cervical cancer in Shandong Province of China. Methods DNA was extracted from cervical cancer biopsy tissue in Shandong Province, China. HPV type was identified by PCR with multiplex PCR and HPV16 type DNA was selected as the template for PCR amplification. HPV16E6E7 gene was obtained and recombined into pALTER -1 vector for bidirectional sequencing and analysis. Results The recombinant plasmid containing HPV16E6E7 in cervical cancer patients was constructed and named as HPV16E6E7-SD. DNA sequence analysis showed that the full length of this sequence was 776bp, which was equal to the length of the published German standard strain, but the nucleotide “T” at nucleotide position 557 of the nucleotide sequence changed to “C”, ie, the stop codon TAA of E6 Change to glutamine codon CAA. Conclusion There is a difference between HPV16E6E7 gene and HPV16E6E7 gene in patients with cervical cancer in Shandong Province of China.