扬州地方株HPV16 L1基因的克隆及序列分析

目的：了解扬州地方株HPV16L1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HPV16L1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HPV16L1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。

Cloning and sequencing of human papillomavirus 16 L1 gene from Yangzhou city

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.