Document Detail

Rapid localization and characterization of random mutations within the 2 micron circle site-specific recombinase: a general strategy for analysis of protein function.
MedLine Citation:
PMID:  2885247     Owner:  NLM     Status:  MEDLINE    
A method for rapidly localizing and characterizing mutations within the 2 micron circle site-specific recombinase enzyme (FLP) is described. The strategy consists of dividing the gene coding for FLP (FLP) by artificially introduced unique restriction enzyme sites (that do not alter the amino acid sequence of the protein) into segments of 150-200 bp, mutagenizing the engineered gene and selecting mutants in vivo, localizing each mutation to one of the gene segments by an in vivo complementation assay, and characterizing the mutation by sequencing of only this DNA segment. The experimental designs embodied by this method should be of wide application in investigations of protein function in general and of DNA-protein interactions in particular.
N S Govind; M Jayaram
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Gene     Volume:  51     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  1987  
Date Detail:
Created Date:  1987-07-24     Completed Date:  1987-07-24     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  31-41     Citation Subset:  IM    
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MeSH Terms
Amino Acid Sequence
Base Sequence
DNA Nucleotidyltransferases / genetics*,  physiology
DNA, Fungal / genetics
Fungal Proteins / genetics*,  physiology
Genes, Synthetic*
Genetic Engineering / methods*
Polymorphism, Restriction Fragment Length
Saccharomyces cerevisiae / genetics
Substrate Specificity
Reg. No./Substance:
0/DNA, Fungal; 0/Fungal Proteins; EC 2.7.7.-/DNA Nucleotidyltransferases; EC 2.7.7.-/FLP recombinase
Erratum In:
Gene 1987;57(1):149

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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