Translation of black beetle virus RNA and heterologous viral RNAs in cell-free lysates derived from Drosophila melanogaster.
Translation of black beetle virus RNA and heterologous viral RNAs in cell-free lysates derived from Drosophila melanogaster.
复制标题
黑甲虫病毒 RNA 和异源病毒 RNA 在黑腹果蝇无细胞裂解物中的翻译。
DOI：
10.1128/jvi.37.1.500-505.1981
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发表时间：
1981
影响因子：
5.4
通讯作者：
Kaesberg,P
中科院分区：
文献类型：
--
作者：
Guarino,LA;Hruby,DE;Ball,LA;Kaesberg,P
由黑腹果蝇1系细胞制备无细胞蛋白质合成系统，并通过微球菌核酸酶处理使其具有mRNA依赖性。使用来自在果蝇细胞中繁殖的黑甲虫病毒的同源RNA、果蝇热休克mRNA以及各种异源病毒mRNA对该系统进行了测试。在最佳条件下，用黑甲虫病毒 RNA 编程的氨基酸掺入比内源掺入高 30 倍。 RNA 1 和 2 主要指导分子量分别约为 120,000 和 46,000 的蛋白质的合成。通过水泡性口炎病毒或牛痘病毒转录制备的 mRNA 被有效翻译并产生与真实病毒蛋白共迁移的产物。雀麦花叶病毒 RNA 和脑心肌炎病毒 RNA 翻译较差。该系统在冷冻后保留了全部活性。
A cell-free protein synthesizing system was prepared from cells of Drosophila melanogaster line 1 and made mRNA dependent by treatment with micrococcal nuclease. The system was tested with homologous RNA from black beetle virus propagated in Drosophila cells, with Drosophila heat shock mRNA, and with various heterologous viral mRNA's. Under optimal conditions amino acid incorporation programmed with black beetle virus RNAs was 30-fold higher than endogenous incorporation. RNAs 1 and 2 primarily directed the synthesis of proteins with approximately molecular weights of 120,000 and 46,000, respectively. mRNA's, prepared by transcription from vesicular stomatitis virus or vaccinia virus, were translated efficiently and yielded products that comigrated with authentic viral proteins. Brome mosaic virus RNA and encephalomyocarditis virus RNA were translated poorly. The system retained full activity after freezing.