Chisholm G E, Henner D J
Department of Cell Genetics, Genentech Inc., South San Francisco, Calfornia 94080.
J Virol. 1988 Sep;62(9):3193-200. doi: 10.1128/JVI.62.9.3193-3200.1988.
The immediate-early IE-1 gene of Autographa californica nuclear polyhedrosis virus was cloned, and its nucleotide sequence was determined. Sequence analysis indicated that this gene would encode a protein of 582 amino acids with a predicted molecular weight of 66,822. Analysis of IE-1 gene expression during baculovirus infection identified two transcripts. One, 1.9 kilobases (kb), was expressed at constant steady-state levels throughout infection, whereas the other, 2.1 kb, was expressed only early in infection. Analysis of IE-1 cDNA clones demonstrated that the 2.1-kb transcript contained the entire 1.9-kb transcript (exon 1) plus an additional 5' end (exon 0). Genomic Southern analysis placed the exon 0 sequences on the EcoRI B fragment, 4 kilobase pairs upstream of exon 1. Sequencing of the upstream region identified an open reading frame whose 5' end was identical to the exon 0 sequences in the cDNAs. Examination of the genomic DNA sequences around the exon-exon junction revealed sequences similar to published consensus splice acceptor and donor sequences. This is the first example of splicing of any viral transcript during baculovirus infection.
克隆了苜蓿银纹夜蛾核型多角体病毒的立即早期IE - 1基因,并测定了其核苷酸序列。序列分析表明,该基因将编码一个由582个氨基酸组成的蛋白质,预测分子量为66,822。对杆状病毒感染期间IE - 1基因表达的分析确定了两种转录本。一种是1.9千碱基(kb),在整个感染过程中以恒定的稳态水平表达,而另一种是2.1 kb,仅在感染早期表达。对IE - 1 cDNA克隆的分析表明,2.1 kb的转录本包含整个1.9 kb的转录本(外显子1)加上一个额外的5'端(外显子0)。基因组Southern分析将外显子0序列定位在EcoRI B片段上,位于外显子1上游4千碱基对处。对上游区域的测序确定了一个开放阅读框,其5'端与cDNA中的外显子0序列相同。对外显子 - 外显子交界处周围的基因组DNA序列的检查揭示了与已发表的共有剪接受体和供体序列相似的序列。这是杆状病毒感染期间任何病毒转录本剪接的第一个例子。