首页> 美国卫生研究院文献>Nucleic Acids Research >Regulation of the herpes simplex virus type 1 late (gamma 2) glycoprotein C gene: sequences between base pairs -34 to +29 control transient expression and responsiveness to transactivation by the products of the immediate early (alpha) 4 and 0 genes.
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Regulation of the herpes simplex virus type 1 late (gamma 2) glycoprotein C gene: sequences between base pairs -34 to +29 control transient expression and responsiveness to transactivation by the products of the immediate early (alpha) 4 and 0 genes.

机译:单纯疱疹病毒1型晚期(γ2)糖蛋白C基因的调控:-34至+29碱基对之间的序列控制瞬时表达和立即早期(alpha)4和0基因产物对反式激活的响应。

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摘要

The glycoprotein C (gC) gene of herpes simplex virus type 1 is a true late gene, in that its expression occurs late in infection with a strict requirement for viral DNA replication. Recently, we reported on gC expression during infection with mutant viruses carrying deletions in the gC gene promoter. Analysis of RNA extracted from cells infected with individual mutants showed that the DNA sequences required for regulated expression of this late gene lie within bases -34 to +124 relative to the 5' end of the mRNA. In the present study, the deleted gC promoter sequences were fused to the bacterial chlorampheniol acetyltransferase (CAT) gene and expression was measured in short-term transfection assays after transactivation by infection with HSV or cotransfection with a second plasmid carrying the immediate early genes of HSV-1. The 63 base pair sequence located between -34 to +29 on the gC promoter was sufficient to give induction of CAT activity following infection and on cotransfection with plasmids which code for the immediate early gene products ICP4 and ICPO. This 63 base pair region contains the TATA homology and the transcriptional start site of the gC gene, and apparently contains specific promoter elements not found in a similar region of the HSV TK promoter. This was shown by removing the distal upstream region of the TK promoter, 5' to -37, and found that the TK gene was no longer activated by infection or cotransfection with an alpha 4-alpha 0 gene containing plasmid.
机译:1型单纯疱疹病毒的糖蛋白C(gC)基因是真正的晚期基因,因为它的表达在感染后期才出现,对病毒DNA复制有严格的要求。最近,我们报道了在携带gC基因启动子缺失的突变病毒感染过程中的gC表达。对从感染了单个突变体的细胞中提取的RNA的分析表明,调节该晚期基因表达所需的DNA序列相对于mRNA的5'端位于-34至+124碱基之间。在本研究中,将缺失的gC启动子序列与细菌氯霉素乙酰基转移酶(CAT)基因融合,并在通过HSV感染进行转激活或与第二个携带HSV早期基因的质粒共转染后,在短期转染测定中测量表达-1。位于gC启动子上的-34至+29之间的63个碱基对的序列足以在感染后以及与编码立即早期基因产物ICP4和ICPO的质粒共转染时诱导CAT活性。该63个碱基对区域包含TATA同源性和gC基因的转录起始位点,并且显然包含在HSV TK启动子的相似区域中未发现的特定启动子元件。这通过除去TK启动子的远端上游区域5'至-37而显示,并且发现TK基因不再被含有α4-α0基因的质粒感染或共转染而激活。

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