本研究以痘苗病毒天坛株(Vaccinia Virus Tian Tan Strain,VTT)为对象,旨在明确N1L基因的表达调控序列.我们克隆了N1L起始密码子ATG上游不同长度的调控序列片段,构建了表达载体,并利用报告基因萤火虫荧光素酶(Firefly luciferase,Fluc)的表达来确定N1L表达调控序列区域.通过PROMO在线预测网站,预测了与调控序列结合的转录因子.结果显示,VTT中N1L基因的表达受到上游调控序列的调控,其中以-404bp起始的片段启动的Fluc表达量最高.PROMO预测表明该区域存在多个转录因子结合位点.本研究为痘苗病毒基因表达特点的研究提供了实验依据,并为N1L基因功能及机制研究奠定了基础.
Cloning and Characteristics of the Regulatory Sequences for N1L Expression in Vaccinia Virus Tian Tan Strain
We aimed to elucidate the regulatory sequences governing expression of N1L in vaccinia virus Tian Tan strain.Various lengths of regulatory sequence fragments upstream of the N1L start codon ATG were cloned,and expression vectors were constructed.Expression of the reporter gene Firefly luciferase(Fluc)served to identify regions of high N1L expression in regulatory sequences.Utilizing an online prediction tool(PROMO),predictions were made regarding transcription factors binding to regulatory sequences.N1L expression in VTT was regulated by upstream regulatory sequences,with the fragment commencing at-404 bp initiating the highest Fluc expression.PROMO predictions indicated the presence of multiple binding sites in transcription factors within this region.This study furnishes experimental evidence for understanding genetic expression in the vaccinia virus,laying the groundwork for further research into the function and mechanism of N1L.
Vaccinia virus Tian Tan strainN1LGene expression and regulationTranscriptional read-through
NETL
NSTL
万方数据
