Esche H, Lübbert H, Siegmann B, Doerfler W
EMBO J. 1982;1(12):1629-33. doi: 10.1002/j.1460-2075.1982.tb01365.x.
We have mapped early and late viral gene products expressed in Autographa californica nuclear polyhedrosis virus ( AcNPV )-infected Spodoptera frugiperda cells by cell-free translation of virus-specific RNA which was selected by hybridization to cloned restriction endonuclease fragments of AcNPV DNA. Proteins synthesized in vitro were labeled with [35S]methionine and analyzed by SDS-polyacrylamide gel electrophoresis followed by fluorography. At least four early AcNPV -specific polypeptides were found which mapped in two regions of the genome (9-25 and 43-59 map units). These early mRNAs are also synthesized at late times in the infection cycle. Cell-free translation of restriction fragment-selected late AcNPV -specific RNA (24 h post-infection) resulted in the identification and mapping of 24 viral proteins. Curiously, the region between approximately 70 and 80 map units on the viral genome has been found silent with respect to mRNA which is translatable in a cell-free system. However, there may be RNA transcribed from this viral DNA segment.
我们通过对病毒特异性RNA进行无细胞翻译,绘制了在感染苜蓿银纹夜蛾核型多角体病毒(AcNPV)的草地贪夜蛾细胞中表达的早期和晚期病毒基因产物图谱。该病毒特异性RNA是通过与AcNPV DNA的克隆限制性内切酶片段杂交筛选出来的。体外合成的蛋白质用[35S]甲硫氨酸标记,并通过SDS-聚丙烯酰胺凝胶电泳和荧光自显影进行分析。发现至少有四种早期AcNPV特异性多肽,它们定位于基因组的两个区域(9 - 25和43 - 59图谱单位)。这些早期mRNA在感染周期的后期也会合成。对感染后24小时的限制性片段筛选的晚期AcNPV特异性RNA进行无细胞翻译,鉴定并绘制了24种病毒蛋白的图谱。奇怪的是,在病毒基因组上大约70至80图谱单位之间的区域,相对于在无细胞系统中可翻译的mRNA而言是沉默的。然而,该病毒DNA片段可能会转录出RNA。