扬州地方株HPV16 L1基因的克隆及序列分析-东南大学学报医学版

扬州地方株HPV16 L1基因的克隆及序列分析

作者：高慧¹ 黄正芳² 徐向明² 孙强² 殷俊² 杨玲² 贾筱琴¹ 朱洪² 李厚达²

单位：1.扬州大学,比较医学中心,江苏,扬州,225009;扬州大学,医学院,江苏,扬州,225001； 2.扬州大学,比较医学中心,江苏,扬州,225009

分类号：Q754

出版年·卷·期（页码）：2005·24·第一期（17-20）

摘要：

目的:了解扬州地方株HPV16-L1基因结构特点,为研制HPV16疫苗奠定基础.方法:从感染HPV16的宫颈癌组织中提取DNA,用PCR技术获得HPV16-L1基因,以pGEM-Teasy为克隆载体构建重组质粒pGEM-T-L1,进行限制性核酸内切酶及核苷酸序列分析.结果:扬州地方株与德国标准株在核苷酸序列上有7处不同,其中4处由于核苷酸的改变,其编码氨基酸也相应发生变化,与标准株的同源性为99.7%;扬州地方株与中国株之间也存在1至2处核苷酸不同,但未造成氨基酸序列改变,其同源性为99.9%.结论:扬州地方株HPV16-L1基因与标准株、中国株之间均存在差异,但与后者的差异极小,未造成氨基酸改变.

Objective To investigate the gene sequence and mutations of HPV16L1 in patients with HPV16 infection in Yangzhou city in order to provide the foundation for preventive vaccine. Methods Sample DNA was extracted from the cervix cancer specimens with HPV16 infection. The HPV16 L1 gene was amplified by PCR and inserted into the pGEMTeasy vector to construct a recombinant plasmid pGEMTeasyL1. Results Restriction endonuclease and sequence analysis showed that there were 7 nucleotide mutation in HPV16L1 compared with the Germanic standard strain. The further amino acid analysis demonstrated that 4 nucleotide mutation induce the changes of amino acid. The sequences of Yangzhou strain were 99.7% homology with Germanic standard strain. There were 1 or 2 nucleotide differences between Yangzhou strain and Chinese strain. There was no change of their amino acids. The sequences were 99.7% homology. Conclusions There are nucleotide differences of HPV16 L1 gene among Germainc standard strain,Chinese strain and Yangzhou strain. Amino acids of Yangzhou strain do not differ from those of Chinese strain.

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