Guttieri M C, Burand J P
Department of Microbiology, University of Massachusetts, Amherst 01003, USA.
Virology. 1996 Sep 15;223(2):370-5. doi: 10.1006/viro.1996.0489.
A late gene of the Hz-1 insect virus (Hz-1V) encoding a predicted polypeptide of 34 kilodaltons (kDa) was isolated from a cDNA library and mapped to the HindIII-T region (50.3 to 52.4 map units) of the viral genome. The p34 gene was characterized by DNA sequence, Northern blot, and primer extension analyses. The 765-bp open reading frame (ORF) is transcribed in the clockwise direction as a 1.2-kb RNA. Primer extension analysis detected two late transcription initiation sites at -16 and -17 nt relative to the start of the p34 ORF. Transcription initiation was observed between 4 and 18 hr postinfection (hr p.i.) with maximum expression at 12 hr p.i. No nucleotide sequence homology was detected between the regulatory region of the p34 gene and the baculovirus conserved late promoter motif NTAAG. This observation was substantiated by results obtained from an investigation of Hz-1V late gene expression using a transient expression assay system which suggested that Hz-1V late gene promoters do not resemble the baculovirus late promoter motif. This is the first molecular analysis of Hz-1V late gene expression and offers a basis by which to compare Hz-1V to other insect viruses.
从cDNA文库中分离出一种编码预测分子量为34千道尔顿(kDa)多肽的Hz - 1昆虫病毒(Hz - 1V)晚期基因,并将其定位到病毒基因组的HindIII - T区域(50.3至52.4个图谱单位)。通过DNA序列分析、Northern印迹分析和引物延伸分析对p34基因进行了表征。765个碱基对的开放阅读框(ORF)以1.2千碱基RNA的形式按顺时针方向转录。引物延伸分析在相对于p34 ORF起始位置-16和-17核苷酸处检测到两个晚期转录起始位点。在感染后4至18小时(hpi)观察到转录起始,在12 hpi时表达量最高。在p34基因的调控区域与杆状病毒保守晚期启动子基序NTAAG之间未检测到核苷酸序列同源性。使用瞬时表达测定系统对Hz - 1V晚期基因表达进行研究所得结果证实了这一观察结果,该研究表明Hz - 1V晚期基因启动子与杆状病毒晚期启动子基序不同。这是对Hz - 1V晚期基因表达的首次分子分析,为将Hz - 1V与其他昆虫病毒进行比较提供了基础。