Document Detail


Effective elimination of nucleic acids from bacterial protein samples for optimized blue native polyacrylamide gel electrophoresis.
MedLine Citation:
PMID:  19598158     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nucleic acids remaining within bacterial protein samples from Streptomyces lividans and Escherichia coli were found to interfere significantly with blue native polyacrylamide gel electrophoresis (BN-PAGE), a technique used frequently for analyzing bacterial protein complexes in proteomics studies. We have used ultracentrifugation and/or precipitation of cell lysates with streptomycin sulfate to eliminate nucleic acids from total and/or membrane protein samples. Nucleic acid-binding proteins were first enriched by precipitation with streptomycin sulfate, and contaminating nucleic acids were then eliminated by precipitation by adding polyethyleneimine. The performance of BN-PAGE was found to be dramatically improved by these sample preparation steps.
Authors:
Jingdan Liang; Qiule Niu; Xinping Xu; Yuanming Luo; Xiufen Zhou; Zixin Deng; Zhijun Wang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Electrophoresis     Volume:  30     ISSN:  1522-2683     ISO Abbreviation:  Electrophoresis     Publication Date:  2009 Jul 
Date Detail:
Created Date:  2009-08-03     Completed Date:  2009-10-27     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  8204476     Medline TA:  Electrophoresis     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  2454-9     Citation Subset:  IM    
Affiliation:
Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, P. R. China.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / chemistry,  isolation & purification*
Chemical Precipitation
DNA, Bacterial / chemistry,  isolation & purification,  metabolism
DNA-Binding Proteins / metabolism
Electrophoresis, Polyacrylamide Gel / methods*
Escherichia coli / chemistry
Escherichia coli Proteins / chemistry
Membrane Proteins / chemistry
Nucleic Acids / chemistry,  isolation & purification*,  metabolism
RNA, Bacterial / chemistry,  isolation & purification,  metabolism
Streptomyces lividans / chemistry
Streptomycin / chemistry
Ultracentrifugation
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/DNA, Bacterial; 0/DNA-Binding Proteins; 0/Escherichia coli Proteins; 0/Membrane Proteins; 0/Nucleic Acids; 0/RNA, Bacterial; 57-92-1/Streptomycin

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