广东地区宫颈癌组织HPV18L1基因的克隆及序列分析

Cloning and sequencing of human papillomavirus type 18L1 gene from cervical cancer tissue in Guangdong area

目的:了解广东地区地方株HPV18L1基因结构特点,为HPV感染的检测和基因工程疫苗的研制提供实验基础。方法:采用PCR技术从广东地区宫颈癌组织中扩增HPV18L1基因,克隆入毕赤酵母表达载体pPICZαB,测序并对HPV18L1基因进行序列分析。结果:广东地区分离株与德国标准株在核苷酸序列上有4处不同,其中3处由于核苷酸的改变,其编码的氨基酸也发生相应变化,与标准株的同源性为99.8%。与中国西安地区分离株比较有两处突变点相同。结论:广东地区HPV18L1分离株与德国标准株、中国其他地区分离株之间均存在差异。

Aim：To clone and analyze the whole sequence of HPV18L1 gene from Guangdong specimen of cervical cancer. Methods： The HPV18L1 gene were amplified from human cervical carcinoma tissues in Guangdong province by PCR, and cloned into Pichia pastories expression vector pPICZaB. The HPV18L1 gene were sequenced and analyzed. Results： The HPV18L1 gene sequence were obtained, and the HPV18L1 Pichia pastories expression vector were constructed. Compare to the standard strain （ German）, there were 4 nucleotide mutations. The further amino acid analysis demonstrated that 3 nucleotide mutations induce the changes of amino acid, the sequences of Guangdong strain were 99.8% homology with Germanic standard strain. Compare to the XiAn strain got by Gao, there were 2 same nucleotide mutations. Conclusion： There are nucleotide differences of the HPV18L1 gene among Germainc standard strain, Gangdong strain and other Chinese strain. Amino acids of Gangdong strain differ from those of Chinese strain.