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目的进一步研究与宫颈癌密切相关的人乳头瘤病毒16型(HPV16)E7基因的生物学活性。方法采用PCR法从中国妇女宫颈癌标本中扩增HPV16早期基因E7,测序分析其DNA的序列。结果克隆的HPV16E7基因与标准型进行比较表明,前者无核苷酸突变。将此基因装入原核表达载体,能在E.Coli中高效表达出谷胱甘肽S-转移酶(GST)-E7蛋白。Western印迹分析表明,此融合蛋白能与E7抗体特异地结合。结论克隆的HPV16E7基因及其表达的蛋白可为今后分子流行病学调查及疫苗研制提供有用的资料。
Objective To further study the biological activity of human papillomavirus type 16 (HPV16) E7 gene, which is closely related to cervical cancer. Methods The HPV16 early gene E7 was amplified from cervical cancer samples of Chinese women by PCR, and its DNA sequence was analyzed by sequencing. Results Cloned HPV16E7 gene compared with the standard type, the former no nucleotide mutation. This gene was inserted into the prokaryotic expression vector and efficiently expressed glutathione S-transferase (GST) -E7 protein in E.coli. Western blot analysis showed that this fusion protein specifically binds to the E7 antibody. Conclusion The cloned HPV16E7 gene and its expressed protein can provide useful information for future molecular epidemiological investigation and vaccine development.