摘要
本文作者采用基因克隆手段,以pAt153为载体,从一份山西襄垣宫颈癌高发区宫颈癌患者的手术标本中,成功地克隆到2株与HPV16同源的基因片段。经PstI、KpnI、TaqvI、PvuII等16种限制性内切酶酶谱分析及其部分基因序列的鉴定,证明这是在国内首次克隆到一侏分子量约为8.0kb完整的HPV16型全序列DNA及一株分子量为5.4kb的HPV16基因片段。经实验证明:该基因片段的E6、E7及部分LI基因丢失,在750单核苷酸处发生变异,产生一新的BamHI酶切位点。该完整的HPV16基因组被命名为HPV16Z,HPV16基因片段被命名为HPV16F。用新分离到的HPV16Z作分子探针,检测襄垣337份宫颈癌及阴道活检标本的HPV16型同源序列的结果显示,慢性阴道炎阳性率为17.28％(14/81);宫颈炎为11.89％(17/143);宫颈癌前病变为46.81%(22/47);宫颈癌为72.73%(48/66)。证明山西宫颈癌高发区宫颈癌前病变及宫颈癌组织中主要为HPV16Z感染。
The cloning and characterization of a whole genome of human Papillomavirus type 16 (7.9kb) and a 5.4kb subgenomic fragment are presented here for the first time in China. Both of them were cloned in HB101 after insertion in plasmid pAt153, using it's unique BamH1 site. The full-length genome and subgenomic fragment are designated HPV16Z and HPV16F, respectively. After analysis of 16 endonucleases, none of new endonuclease site were found in HPV16Z comparing with standard strain. A new BamH1 site was observed in 750bp in HPV16F. DNA sequencing will be done in next experiment. Using HPV16Z as molecular probe, 337 cervix and vagina biopsies from Xiang Yuan County were tested by means of Coslot hybridization at high strict condition (Tm-17℃). The experiment result showed that the HPV16Z infectious rates of chronic cervicitis, vaginitis, cervical pre-cancer lesions (CIN I-CIN Ⅲ), cervical cancer are 11.89% (17/143), 17.28% (14/81), 46.81%(22/47) and 72.73(48/66), respectively. The above information indicated that HPV16Z infection is predominant in the cervix pre-cancer lesion and cervical cancer at the cervical cancer high incidence region.
出处
《中国病毒学》
CSCD
1993年第1期45-52,共8页
Virologica Sinica
基金
该项研究为国家"七五"资助课题
