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在由市售小麦胚芽制备的无细胞体系中烟草花叶病毒RNA和兔珠蛋白9S RNA的高效翻译。

Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ.

作者信息

Roberts B E, Paterson B M

出版信息

Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330-4. doi: 10.1073/pnas.70.8.2330.

DOI:10.1073/pnas.70.8.2330
PMID:4525168
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC433729/
Abstract

Extracts prepared from commercially available wheat germ efficiently translate messenger RNAs from either viral or eukaryotic origin. The addition of tobacco mosaic virus RNA stimulated amino-acid incorporation more than 100-times and rabbit globin 9S RNA between 20- and 30-times. The in vitro product directed by globin 9S RNA comigrated precisely with authentic rabbit globin on sodium dodecyl sulfate-polyacrylamide gels. Furthermore, during high voltage ionophoresis two [(35)S]methionine-labeled tryptic peptides synthesized in vitro comigrated in two dimensions with alphaT5 and betaT5 tryptic peptides from authentic [(35)S]methionine-labeled rabbit globin.

摘要

从市售小麦胚芽中提取的提取物能够高效翻译病毒或真核生物来源的信使核糖核酸。添加烟草花叶病毒核糖核酸可使氨基酸掺入量增加100倍以上，添加兔珠蛋白9S核糖核酸可使氨基酸掺入量增加20至30倍。在十二烷基硫酸钠-聚丙烯酰胺凝胶上，由珠蛋白9S核糖核酸指导的体外产物与天然兔珠蛋白精确共迁移。此外，在高压离子电泳过程中，体外合成的两种[(35)S]甲硫氨酸标记的胰蛋白酶肽在二维上与天然[(35)S]甲硫氨酸标记的兔珠蛋白的αT5和βT5胰蛋白酶肽共迁移。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e072/433729/69898b12de29/pnas00071-0151-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e072/433729/4c3a5d5bd941/pnas00071-0150-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e072/433729/69898b12de29/pnas00071-0151-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e072/433729/4c3a5d5bd941/pnas00071-0150-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e072/433729/69898b12de29/pnas00071-0151-a.jpg

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1
Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ.在由市售小麦胚芽制备的无细胞体系中烟草花叶病毒RNA和兔珠蛋白9S RNA的高效翻译。
Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330-4. doi: 10.1073/pnas.70.8.2330.
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Cell-Free Gene Expression: Methods and Applications.无细胞基因表达：方法与应用
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Advancing synthetic biology through cell-free protein synthesis.通过无细胞蛋白质合成推动合成生物学发展。
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Cell-Free Expression of Designed Peptides That Form β-Barrel Nanopores.无细胞表达设计肽形成β-桶纳米孔。
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Development of a cell-free protein synthesis system for practical use.无细胞蛋白质合成系统的开发及其实际应用。
Proc Jpn Acad Ser B Phys Biol Sci. 2021;97(5):261-276. doi: 10.2183/pjab.97.015.
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Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.复杂生物分子组装体的简易合成：结构生物学中的小麦胚无细胞蛋白质表达
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