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微红盘绒茧蜂(膜翅目:茧蜂科)一种多分DNA病毒基因的启动子研究

Promoter studies of a polydnavirus gene from Cotesia rubecula (Hym: Braconidae).

作者信息

Asgari S, Schmidt O

机构信息

Department of Applied and Molecular Ecology, Adelaide University, Glen Osmond, Australia.

出版信息

Arch Virol. 2001 Oct;146(10):1979-89. doi: 10.1007/s007050170046.

Abstract

The Cotesia rubecula polydnavirus gene, CrV1, is expressed in a highly transient fashion. Within four hours after egg deposition and virus infection, tissues of the host caterpillar, Pieris rapae, express high levels of the transcript. Twelve hours after infection no transcripts are visible. We have previously shown that the CrV1 secreted protein is mainly produced in host haemocytes. In haemocytes, immune functions such as phagocytosis and cell spreading are abolished by destabilization of the cell cytoskeleton. To test whether the observed down-regulation of CrV1 transcripts is mediated by transcriptional control or by other factors, such as the disruption of cytoskeleton in CrV1-inactivated cells, we cloned the promoter and the 3' untranslated region of the CrV1 gene to study CrV1 expression. The promoter region of the CrV1 gene was cloned into baculovirus expression systems along with the CAT reporter gene. Molecular analyses showed that the CAT gene under the control of CrV1 promoter is expressed as early as 2 h post infection and continues until late phase of infection suggesting that down-regulation of CrV1 expression in host haemocytes is perhaps mediated by post-transcriptional mechanisms.

摘要

菜蛾盘绒茧蜂多DNA病毒基因CrV1以高度短暂的方式表达。在产卵和病毒感染后四小时内,宿主菜粉蝶毛虫的组织会高水平表达该转录本。感染十二小时后则看不到转录本。我们之前已经表明,CrV1分泌蛋白主要在宿主血细胞中产生。在血细胞中,吞噬作用和细胞铺展等免疫功能会因细胞骨架的不稳定而被消除。为了测试观察到的CrV1转录本下调是由转录控制还是由其他因素介导的,例如CrV1失活细胞中细胞骨架的破坏,我们克隆了CrV1基因的启动子和3'非翻译区来研究CrV1的表达。CrV1基因的启动子区域与CAT报告基因一起被克隆到杆状病毒表达系统中。分子分析表明,在CrV1启动子控制下的CAT基因在感染后2小时就开始表达,并持续到感染后期,这表明宿主血细胞中CrV1表达的下调可能是由转录后机制介导的。

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