دورية أكاديمية
Purification of 30S ribosomal subunit by streptavidin affinity chromatography.
| العنوان: | Purification of 30S ribosomal subunit by streptavidin affinity chromatography. |
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| المؤلفون: | Golovina AY; Department of Chemistry, Moscow State University, Moscow 119992, Russia., Bogdanov AA, Dontsova OA, Sergiev PV |
| المصدر: | Biochimie [Biochimie] 2010 Jul; Vol. 92 (7), pp. 914-7. Date of Electronic Publication: 2010 Mar 25. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Editions Scientifiques Elsevier Country of Publication: France NLM ID: 1264604 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1638-6183 (Electronic) Linking ISSN: 03009084 NLM ISO Abbreviation: Biochimie Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Paris : Editions Scientifiques Elsevier Original Publication: Paris. |
| مواضيع طبية MeSH: | Chromatography, Affinity/*methods , Ribosome Subunits, Small, Bacterial/*metabolism , Streptavidin/*metabolism, Aptamers, Nucleotide/genetics ; Aptamers, Nucleotide/metabolism ; Base Sequence ; Escherichia coli/cytology ; Molecular Sequence Data ; Nucleic Acid Conformation ; Ribosome Subunits, Small, Bacterial/chemistry |
| مستخلص: | Preparation of pure ribosomal subunits carrying lethal mutations is necessary for studying every essential functional region of ribosomal RNA. Affinity purification via a tag, inserted into rRNA proved to be procedure of choice for purification of such ribosomal subunits. Here we describe fast and simple purification method for the 30S ribosomal subunits using affinity chromatography. Streptavidin-binding tag was inserted into functionally neutral helix 33a of the 16 S rRNA from Escherichia coli. Tagged ribosomal subunits were shown to be expressed in E. coli and could be purified. Purified subunits with affinity tag behave similarly to the wild type subunits in association with the 50S subunits, toe-printing and tRNA binding assays. Tagged 30S subunits could support cell growth in the strain lacking wild type 30S subunits and only marginally change the growth rate of bacteria. The presented purification method is thus suitable for further use in purification of 30S subunits carrying any lethal mutations. (Copyright 2010 Elsevier Masson SAS. All rights reserved.) |
| المشرفين على المادة: | 0 (Aptamers, Nucleotide) 9013-20-1 (Streptavidin) |
| تواريخ الأحداث: | Date Created: 20100330 Date Completed: 20100924 Latest Revision: 20100623 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1016/j.biochi.2010.03.012 |
| PMID: | 20347003 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1638-6183 |
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| DOI: | 10.1016/j.biochi.2010.03.012 |