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利用哺乳动物无细胞提取物对心病毒核糖核酸进行体外翻译。

In vitro translation of cardiovirus ribonucleic acid by mammalian cell-free extracts.

作者信息

Eggen K L, Shatkin A J

出版信息

J Virol. 1972 Apr;9(4):636-45. doi: 10.1128/JVI.9.4.636-645.1972.

Abstract

Cell-free extracts prepared from Ehrlich ascites and mouse L cells synthesize viral proteins in response to encephalomyocarditis virus, mouse Elberfeld virus, and mengovirus ribonucleic acid. Although HeLa cell extracts are inactive, their ribosomes are functional in the presence of heterologous supernatant fractions. Synthesis depends upon the addition of adenosine triphosphate, guanosine triphosphate, an energy-generating system, and 4 mm Mg(2+). Initiation is completed during the first 10 to 20 min of incubation, but chain elongation continues for 1 hr or more. The products are of higher molecular weight than virion structural proteins and resemble polypeptides formed in virus-infected cells during a short pulse. Tryptic peptides of virion proteins and in vitro products are similar for all three cardioviruses.

摘要

从艾氏腹水瘤细胞和小鼠L细胞制备的无细胞提取物,能响应脑心肌炎病毒、小鼠埃尔伯费尔德病毒和门戈病毒核糖核酸合成病毒蛋白。虽然HeLa细胞提取物无活性,但其核糖体在存在异源上清液组分时具有功能。合成依赖于添加三磷酸腺苷、三磷酸鸟苷、能量产生系统和4 mM镁离子。起始反应在孵育的最初10至20分钟内完成,但链延伸持续1小时或更长时间。产物的分子量高于病毒粒子结构蛋白,类似于在病毒感染细胞中短脉冲期间形成的多肽。所有三种心脏病毒的病毒粒子蛋白和体外产物的胰蛋白酶肽相似。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b251/356348/892ea0965637/jvirol00268-0086-a.jpg

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