Lu M, Iatrou K
Department of Medical Biochemistry, The University of Calgary, Alberta, Canada.
J Gen Virol. 1996 Dec;77 ( Pt 12):3135-43. doi: 10.1099/0022-1317-77-12-3135.
We have cloned and analysed the transcriptional properties of two closely linked genes, p39 and cg30, of Bombyx mori nuclear polyhedrosis virus (BmNPV). These genes encode a structural polypeptide and a putative transcriptional regulator of the virus, respectively. The cg30 gene is transcribed prior to and after DNA replication from a site located within the ORF of the adjacent p39 gene. Its transcription product, a 1.3 kb mRNA, is polyadenylated at a site containing consensus eukaryotic polyadenylation signals and mapping 87 bp downstream of the translation termination codon for CG30. During the later stages of infection, two additional RNAs, 2.2 and 6.5 kb, are also transcribed through the cg30 gene. The 2.2 kb RNA, representing the mRNA that encodes P39, is initiated from three relatively closely spaced sites located upstream of the P39 ORF. The 6.5 kb RNA is apparently transcribed from the promoter sequences of another gene located further upstream of the p39 gene. The 2.2 and 6.5 kb transcripts have two polyadenylation sites. The first site is the same as the one used to generate the cg30 gene-specific transcripts. The second is located 4 bp downstream of the CG30 translation termination codon. Transient expression assays show that the p39 gene sequences immediately upstream of the CG30 ORF can direct expression of a reporter gene when the latter is co-transfected with the gene encoding the early baculovirus trans-activator IE1. Thus, these sequences behave as a delayed-early baculovirus gene promoter.
我们克隆并分析了家蚕核型多角体病毒(BmNPV)两个紧密相连的基因p39和cg30的转录特性。这些基因分别编码病毒的一种结构多肽和一种假定的转录调节因子。cg30基因在DNA复制之前和之后从相邻p39基因开放阅读框内的一个位点转录。其转录产物,一个1.3 kb的mRNA,在一个含有真核生物多聚腺苷酸化信号共有序列的位点进行多聚腺苷酸化,该位点位于CG30翻译终止密码子下游87 bp处。在感染后期,另外两种RNA,2.2 kb和6.5 kb,也通过cg30基因转录。2.2 kb的RNA代表编码P39的mRNA,从位于P39开放阅读框上游三个相对紧密间隔的位点起始。6.5 kb的RNA显然是从位于p39基因上游更远位置的另一个基因的启动子序列转录而来。2.2 kb和6.5 kb的转录本有两个多聚腺苷酸化位点。第一个位点与用于产生cg30基因特异性转录本的位点相同。第二个位点位于CG30翻译终止密码子下游4 bp处。瞬时表达分析表明,当CG30开放阅读框紧邻的上游p39基因序列与编码早期杆状病毒反式激活因子IE1的基因共转染时,能够指导报告基因的表达。因此,这些序列具有延迟早期杆状病毒基因启动子的作用。