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甲型流感病毒RNA聚合酶三个亚基的纯化、硫氧还蛋白复性及重组活性

Purification, thioredoxin renaturation, and reconstituted activity of the three subunits of the influenza A virus RNA polymerase.

作者信息

Szewczyk B, Laver W G, Summers D F

机构信息

Department of Cellular, Viral, and Molecular Biology, University of Utah School of Medicine, Salt Lake City 84132.

出版信息

Proc Natl Acad Sci U S A. 1988 Nov;85(21):7907-11. doi: 10.1073/pnas.85.21.7907.

DOI:10.1073/pnas.85.21.7907
PMID:3054875
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC282313/
Abstract

The virion-associated RNA polymerase and the structural nucleoprotein of influenza A virus were separated by sodium dodecyl sulfate/PAGE, electroblotted to a polyvinylidine membrane, and eluted with good recovery from the membrane. After renaturation by incubating with Escherichia coli thioredoxin, these proteins were active in a reconstituted in vitro transcription reaction with purified genomic RNAs. All four proteins (i.e., the three subunits of the RNA polymerase as well as the structural nucleoprotein) were required for activity. The RNA products were plus-strand, mRNA-sized species.

摘要

甲型流感病毒的病毒粒子相关RNA聚合酶和结构核蛋白通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳分离，电印迹到聚偏二氟乙烯膜上，并从膜上洗脱，回收率良好。与大肠杆菌硫氧还蛋白一起孵育复性后，这些蛋白质在与纯化的基因组RNA的体外转录反应重建中具有活性。活性需要所有四种蛋白质（即RNA聚合酶的三个亚基以及结构核蛋白）。RNA产物为正链、mRNA大小的物种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/389992ca1833/pnas00300-0089-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/fa9e7d770153/pnas00300-0087-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/6c91a24e7a37/pnas00300-0088-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/389992ca1833/pnas00300-0089-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/fa9e7d770153/pnas00300-0087-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/6c91a24e7a37/pnas00300-0088-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f53/282313/389992ca1833/pnas00300-0089-a.jpg

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本文引用的文献

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Characterization of influenza virus RNA complete transcripts.流感病毒RNA完整转录本的特征分析
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Influenza virus PB1 protein is the minimal and essential subunit of RNA polymerase.流感病毒PB1蛋白是RNA聚合酶的最小必需亚基。
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