摘要
为研究ＨＰＶ１６转录调节蛋白ＹＹ１结合位点改变对病毒致癌性的影响，以ＨＰＶ１６野毒株质粒ｐ１２０３为基础，经多次克隆，将来自宫颈癌组织并带有缺损突变的ＬＣＲ重组到地ＨＰＶ１６基因组中。核酸序列分析证实，质粒ｐＤＶ３９０在第２个ＹＹ１位点上有一Ｇ→Ａ点穷变，质粒ｐＤＶ３２６和ｐＤＶ４０１分别带有１１５ｂｐ和１４３ｂｐ的缺失突变，从而涉及２和４个ＹＹ１结合位点，重组质粒其它核苷酸序列与ＨＰＶ１６标准序列一致。这些重组质粒的组建为进一步研究ＹＹ１蛋白对ＨＰＶ１６致癌基因表达的调控及ＨＰＶ１６致癌作用的影响打下基础。
Y1 is an imporiant cellular transcriptional regulator which is ubiquitously expressed in vari-ous kinds of cells.The specific binding sites of YY1 protein are found in the flanking sequences ofnlany cellular aiid viral promoters.In the 'high risk'huinan papilloniaviruses(HPVs)YY1 works as a traxiscriptional repressor for the viral early gene promoter.In the episomal HPV 16 genomesextracted from cervieal cancers,removal of YY1 binding sites in LCR is a repeated event, resultingin increasing the activity of viral oncogene promoter P97.In order to study the effect of revomal ofYY1 binding sites in LCR on the viral tumorigenicity,HPV16 wild-tny plasmid p1203 was usedas basic sequences,through mltiple-step-clone to construct the new HPV16 plasnuds containingdeleted and mutated LCRs. houence analysis coofirmed that pDV390 had a G to A exchiinge inthe second YY1 site,whereas PDV 1326 and pDV401 contained 115 bp aiid 143 bp deletions in therespective LCRs that revomed 2 and 4 YY1 binding sites.The rest nucleotide sequences of thenew plasmids showed no variation comparing with HPV 16 wild-tape sequence.These reconstrnct-ed HPV 16 genome plasmids supply a basic work for study of YYI protein on the regulation forthe expression of viral oncogenes as well as viral oncogenicity.
出处
《中国病毒学》
CSCD
1996年第3期237-243,共7页
Virologica Sinica
