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To study trans-activation of gene expression by murine cytomegalovirus (MCMV) immediate-early (IE) proteins, the IE coding region 1 (ie1), which encodes the 89,000-Mr IE phosphoprotein (pp89), was stably introduced into L cells. A cell line was selected and characterized that efficiently expressed the authentic viral protein. The pp89 that was constitutively expressed in L cells stimulated the expression of transfected recombinant constructs containing the bacterial chloramphenicol acetyltransferase (CAT) gene under the control of viral promoters. The regulatory function of the ie1 product was confirmed by transient expression assays in which MCMV IE genes were cotransfected into L cells together with recombinant constructs of the CAT gene. For CAT activation by the ie1 product, a promoter region was required, but there was no preferential activation of a herpes simplex virus type 1 delayed-early promoter. All plasmid constructs that contained the intact coding sequences for pp89 induced gene expression in trans. The MCMV enhancer region was not essential for the expression of a functional IE gene product, and testing of the cis-regulatory activity of the MCMV enhancer revealed a low activity in L cells. Another region transcribed at IE times of infection, IE coding region 2, was unable to induce CAT expression and also did not augment the functional activity of ie1 after cotransfection.  相似文献   

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长非编码RNA研究进展   总被引:2,自引:0,他引:2  
长非编码RNA是指一类长度大于200个核苷酸、不编码蛋白质的非编码RNA.越来越多的研究表明,人类基因组中高达90%的非编码蛋白质的区段同样具有重要作用,而不是所谓的"转录噪声".针对长非编码RNA的功能研究表明,其在转录起始的调控、转录及转录后的调控中均发挥着重要作用,因而影响着各种各样的生物学过程.本综述围绕近几年长非编码RNA的研究成果,总结了长非编码RNA的起源与进化、新型的长非编码RNA类型、典型的长非编码RNA作用机制以及长非编码RNA在发育与细胞重编程过程中的研究,同时也概述了长非编码RNA与表观遗传调控和癌症的关系以及长非编码RNA研究的相关技术.系统发现长非编码RNA并阐明其功能机制,将对现代生命科学具有重大的意义.  相似文献   

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