O'Reilly D R, Passarelli A L, Goldman I F, Miller L K
Department of Entomology, University of Georgia, Athens 30602.
J Gen Virol. 1990 May;71 ( Pt 5):1029-37. doi: 10.1099/0022-1317-71-5-1029.
A region of the baculovirus Autographa californica nuclear polyhedrosis virus genome that is frequently found to be altered after serial passage of the virus in cell culture was characterized. Sequence analysis of this region of the genome in wild-type and mutant viruses revealed that some of the mutations affected a 675 bp open reading frame, designated DA26. The DA26 gene was disrupted both by deletion and by insertion of sequences that resembled transposable elements. Northern blot analysis of DA26 showed that it was expressed very early after infection. DA26-specific transcripts could be detected after the 1 h viral adsorption period upon infection of cultured Trichoplusia ni cells. These transcripts were mapped by nuclease protection assays. A recombinant virus was constructed in which DA26 was disrupted by insertion of the Escherichia coli lacZ gene. This virus was viable in both T. ni and Spodoptera frugiperda cells and analysis of the kinetics of protein synthesis revealed no differences between wild-type and recombinant viruses. The disruption of DA26 also did not interfere with the ability of the virus to infect T. ni or S. frugiperda larvae.
对杆状病毒苜蓿银纹夜蛾核型多角体病毒基因组的一个区域进行了表征，该区域在病毒于细胞培养物中连续传代后经常发生改变。对野生型和突变型病毒基因组的该区域进行序列分析发现，一些突变影响了一个675 bp的开放阅读框，命名为DA26。DA26基因因类似于转座元件的序列的缺失和插入而被破坏。对DA26的Northern印迹分析表明，它在感染后很早便开始表达。在培养的粉纹夜蛾细胞感染后1小时的病毒吸附期过后，即可检测到DA26特异性转录本。这些转录本通过核酸酶保护试验进行了定位。构建了一种重组病毒，其中DA26因插入大肠杆菌lacZ基因而被破坏。该病毒在粉纹夜蛾和草地贪夜蛾细胞中均能存活，对蛋白质合成动力学的分析表明，野生型病毒和重组病毒之间没有差异。DA26的破坏也不影响病毒感染粉纹夜蛾或草地贪夜蛾幼虫的能力。
