Document Detail


Auto-induction medium containing glyphosate for high-level incorporation of unusual aromatic amino acids into proteins.
MedLine Citation:
PMID:  20854268     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe the use of an auto-induction medium containing N-(phosphono-methyl)glycine (glyphosate) as a means for high-level introduction of nonstandard aromatic amino acids into a protein. We illustrate this approach by preparing maltose binding protein (MBP) wherein all eight tryptophan residues have been replaced with 6-fluorotryptophan at an incorporation level of 99.3%. Such a high level of incorporation is important for spectroscopic investigations, in particular 19F NMR, because each species' differing amino acid sequence potentially yields a different peak pattern that complicates spectral analysis.
Authors:
Muniasamy Neerathilingam; John L Markley
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  BioTechniques     Volume:  49     ISSN:  1940-9818     ISO Abbreviation:  BioTechniques     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-09-21     Completed Date:  2011-01-19     Revised Date:  2011-07-20    
Medline Journal Info:
Nlm Unique ID:  8306785     Medline TA:  Biotechniques     Country:  England    
Other Details:
Languages:  eng     Pagination:  659-61     Citation Subset:  IM    
Affiliation:
Biochemistry Department, University of Wisconsin-Madison, Madison, WI 53706, USA.
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MeSH Terms
Descriptor/Qualifier:
Biotechnology / methods
Escherichia coli / genetics,  metabolism*
Glycine / analogs & derivatives*,  metabolism
Herbicides / metabolism*
Maltose-Binding Proteins / chemistry,  genetics,  metabolism*
Recombinant Proteins / chemistry,  genetics,  metabolism
Tryptophan / analogs & derivatives*,  chemistry,  genetics,  metabolism
Grant Support
ID/Acronym/Agency:
1U54 GM074901/GM/NIGMS NIH HHS; P41 RR002301-15/RR/NCRR NIH HHS; P41 RR002301-15S1/RR/NCRR NIH HHS; P41 RR002301-16/RR/NCRR NIH HHS; P41 RR002301-17/RR/NCRR NIH HHS; P41 RR002301-18/RR/NCRR NIH HHS; P41 RR002301-19/RR/NCRR NIH HHS; P41 RR002301-20/RR/NCRR NIH HHS; P41 RR002301-21A1/RR/NCRR NIH HHS; P41 RR002301-22/RR/NCRR NIH HHS; P41 RR002301-23/RR/NCRR NIH HHS; P41 RR002301-23S1/RR/NCRR NIH HHS; P41 RR002301-24/RR/NCRR NIH HHS; P41 RR002301-25/RR/NCRR NIH HHS; P41 RR002301-25S1/RR/NCRR NIH HHS; P41 RR002301-26/RR/NCRR NIH HHS; P50 GM064598-01/GM/NIGMS NIH HHS; P50 GM064598-01S1/GM/NIGMS NIH HHS; P50 GM064598-02/GM/NIGMS NIH HHS; P50 GM064598-03/GM/NIGMS NIH HHS; P50 GM064598-03S1/GM/NIGMS NIH HHS; P50 GM064598-04/GM/NIGMS NIH HHS; P50 GM064598-04S1/GM/NIGMS NIH HHS; P50 GM64598/GM/NIGMS NIH HHS; RR02301/RR/NCRR NIH HHS; U54 GM074901-01/GM/NIGMS NIH HHS; U54 GM074901-01S1/GM/NIGMS NIH HHS; U54 GM074901-02/GM/NIGMS NIH HHS; U54 GM074901-02S1/GM/NIGMS NIH HHS; U54 GM074901-03/GM/NIGMS NIH HHS; U54 GM074901-03S1/GM/NIGMS NIH HHS; U54 GM074901-04/GM/NIGMS NIH HHS; U54 GM074901-05/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Herbicides; 0/Maltose-Binding Proteins; 0/Recombinant Proteins; 1071-83-6/glyphosate; 343-92-0/6-fluorotryptophan; 56-40-6/Glycine; 73-22-3/Tryptophan
Comments/Corrections

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