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利用小麦胚芽体外系统对卫星烟草坏死病毒核糖核酸进行的翻译。

Translation of satellite tobacco necrosis virus ribonucleic acid by an in vitro system from wheat germ.

作者信息

Leung D W, Gilbert C W, Smith R E, Sasavage N L, Clark J M

出版信息

Biochemistry. 1976 Nov 2;15(22):4943-50. doi: 10.1021/bi00667a030.

Abstract

The RNA of satellite tobacco necrosis virus (STNV) is an effective messenger RNA when translated in an in vitro system from wheat germ. This RNA codes for only STNV coat protein, as indicated (1) by coincidence of the tryptic fingerprints of the translation product and of STNV coat protein, (2) by equivalent size of the translation product and STNV coat protein, and (3) by isolation of an initial peptide of the in vitro product containing the amino acid sequence of the N terminus of STNV coat protein. STNV RNA does not contain a 5'-terminal m7G(5')ppp(5')Np---group and translation of STNV RNA by the wheat germ system does not involve prior formation of 5'-terminal m7G(5')ppp(5') nP---groups on STNV RNA. STNV RNA and 125I-labeled STNV RNA form a specific initiation complex when incubated with initiator tRNA, GTP, initiation factors, and wheat germ ribosomes. Treatment of this specific initiation complex with ribonuclease A allows isolation of an 125I-labeled oligonucleotide protected from ribonuclease A by the initiation complex. This specific oligonucleotide contains approximately 38 nucleotides, including nucleotide sequences that coincide with the codons of the N-terminal amino acids of STNV coat proteins.

摘要

卫星烟草坏死病毒(STNV)的RNA在小麦胚芽体外系统中进行翻译时是一种有效的信使RNA。这种RNA仅编码STNV外壳蛋白,这一点可通过以下几点得到证明:(1)翻译产物和STNV外壳蛋白的胰蛋白酶指纹图谱一致;(2)翻译产物和STNV外壳蛋白大小相当;(3)分离出体外产物的起始肽,其包含STNV外壳蛋白N端的氨基酸序列。STNV RNA不含有5'-末端m7G(5')ppp(5')Np---基团,并且小麦胚芽系统对STNV RNA的翻译不涉及在STNV RNA上预先形成5'-末端m7G(5')ppp(5') nP---基团。当与起始tRNA、GTP、起始因子和小麦胚芽核糖体一起孵育时,STNV RNA和125I标记的STNV RNA会形成特异性起始复合物。用核糖核酸酶A处理这种特异性起始复合物,可以分离出一种被起始复合物保护而免受核糖核酸酶A作用的125I标记的寡核苷酸。这种特异性寡核苷酸包含大约38个核苷酸,包括与STNV外壳蛋白N端氨基酸密码子一致的核苷酸序列。

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