天津科技大学学报2024，Vol.39Issue(4)：9-14,25,7.DOI:10.13364/j.issn.1672-6510.20230167
HPV18间隔序列介导真核基因以双顺反子形式表达
HPV18 Intergenic Spacer Mediating Bicistronic Expression in Eukaryotes
摘要
Abstract
HPV18 is a DNA virus,with E6 and E7 gene open reading frame(ORF)separated by 8 nucleotides and expressed as a bicistron in cervical cancer HeLa cells.To investigate whether this 8 nt spacer sequence plays a crucial role in the bicis-tron gene expression,two bicistron expressing plasmids were constructed using this spacer sequence and transfected into HEK293T cells to detect the expression of both upstream and downstream ORFs.RT-PCR results showed simultaneous tran-scription of two ORFs;Western blot results showed that both proteins were expressed simultaneously.The confocal micro-scopy results further demonstrated the co-expression of red fluorescence protein(RFP)and green fluorescence protein(GFP).The above results demonstrate that the HPV18 E6 and E7 gene spacer sequence can re-initiate downstream ORF translation after the completion of upstream ORF translation.This phenomenon is relatively rare in eukaryotic cells,and the molecular mechanism might be related to the reentry of ribosomes which is worthy of further exploration.关键词
翻译起始/多顺反子/HPV/基因表达/核糖体结合序列/真核细胞Key words
translation initiation/polycistron/HPV/gene expression/ribosome binding sequence/eukaryocyte分类
医药卫生引用本文复制引用
张煜豪,孙小强,鲍正好,李雨彤,刘佳艺,何红鹏..HPV18间隔序列介导真核基因以双顺反子形式表达[J].天津科技大学学报,2024,39(4):9-14,25,7.基金项目
国家重点研发计划资助项目(2018YFA0901702) （2018YFA0901702）
天津市级大学生创新创业训练计划资助项目(202310057267) （202310057267）
