不同地区HPV16 E7基因的克隆及序列差异分析

Cloning,Sequencing and Comparing Analysis of Human Papillomavirus Type 16 E7 Gene Isolated from Different Region

采用PCR扩增、pGEM T载体克隆和核苷酸序列分析的方法对一例武汉地区及两例五峰县高发区宫颈癌患者体内HPV16型的E7基因编码区进行序列分析并与野生型 (德国标准株 )及已发表的HPV16湖北株 (HPVHB)进行了比较。结果发现武汉地区HPV16型E7基因仅第 5 4位出现一个同义突变 ,而高发区HPV16型E7基因存在差异 ,第 77位氨基酸由精氨酸 (Arg)变为半胱氨酸 (Cys) ,第 96位由谷氨酰氨酸 (Gln)变为精氨酸 (Arg) ,E7蛋白的二级结构及亲、疏水性也相应改变 。

In order to study the structure specificity of HPV16 E 7 gene of cervical carcinoma in Wuhan city and Wufeng county of Hubei the tissue DNA was abstracted from cervical carcinoma biopsies from Wuhan and high incidence region of cervical carcinoma in Wuhan county.HPV16 E7 gene was amplified by PCR from the cervical carcinoma tissus DNA with the infection of HPV16,and then was cloned into pGEM T vector.After sequencing the double strands,the gene was compared with the prototype E 7 gene of HPV16,HPV16 Hubei strain and compared with each other.Sequencing results showed one point mutation in the viral neocleotide sequence of high incidence region that the 229th neocleotide 'C' was changed into 'T',which caused the Arg cordon TGT to convert into a Cys cordon CGT.This may be the cause of high incidence of HPV infection and high oncogenisis.The results also show that there is only a synonymous mutation in the viral neocleotide sequence of Wuhan which is different from the HPV16 HB E 7 gene,suggesting that HPV16 HB and the HPV16 prototype are co exist in Hubei,but the HPV16 prototype are the main type.