摘要
构建含人乳头瘤病毒11型(Human papillomavirus type11,HPV11)完整开放读码框(Open reading frame,ORF)基因组的质粒并对其功能进行初步验证,为构建HPV11转基因动物模型奠定基础。分别构建重组质粒pQE-Trisystem-EGFP/HPV11(pE/H)、pQE-Trisystem-EGFP/1.1copyHPV11(pE/1.1H),将pE/1.1H、pE/H、闭环状HPV11基因组分别转染原代人角质形成细胞(Keratinocyte,KC)并进行检测。在质粒上成功构建了目的序列,转染后在pE/1.1H组、pE/H组、HPV11组中检测到HPV11E6基因的表达;在pE/1.1H组、pE/H组中检测到荧光;HPV11组、pE/H组、pE/1.1H组具备促细胞增殖功能,实验组间比较pE/1.1H组稍弱(P<0.01),但与对照组相比均差异极显著(P<0.01)。将具有完整ORF的HPV11基因组(1.1copyHPV11)成功构建于质粒上,其具备野生型HPV11病理表型;为研究低危型HPV致病机理提供了实验材料和方法学指导;为进一步构建HPV11转基因小鼠奠定了基础。
We constructed the plasmid containing the integrated open reading frame （ORF） HPV11 genome, and identified its function, to lay foundation for production of transgenic animal models of HPV 11. Recombinant plasmid pQE-Trisystem- EGFP/HPV11 （pE/H） and pQE-Trisystem-EGFP/1.1 copy HPV11 （pE/1.1H） were constructed. Then, the human keratinocyte （KC） was transfected by pE/1.1H, pE/H and closed circular HPV11 and detected. The recombinant plasmid was successfully constructed. The expression of HPV11E6 gene was detected in the experimental group. Fluorescence was observed in the pE/1.1H and pE/H group. The HPV11, pE/H, and pE/1.1 enhanced the KC proliferation, with remarkable differences （P〈0.01） from the control group. Amongst the three experimental groups, pE/1.1H was found to be the weakest. The plasmid containing the integrated ORF of HPV11 （1.1 copy HPV11） genome was successfully constructed. The pE/1.1H had the same phenotype of wild-type HPV11 genome. It provided experimental material and methodological guidance for studying the low-risk HPV, as well as for the production of HPV 11 transgenic mice.
出处
《生物工程学报》
CAS
CSCD
北大核心
2008年第8期1367-1372,共6页
Chinese Journal of Biotechnology
基金
国家自然科学基金项目(No.30400386)资助~~
