Document Detail


Gene trapping with GFP: the isolation of developmental mutants in the slime mold Polysphondylium.
MedLine Citation:
PMID:  9382807     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In order to study how a cell mass undergoes a transition from one symmetry to another in the slime mold Polysphondylium, we developed a genetic screen in which mutant phenotype and gene expression can easily be visualized in the living organism. The screen combines restriction enzyme-mediated integration (REMI) [1,2] and green fluorescent protein (GFP) [3] expression. In REMI, a restriction enzyme is electroporated along with linearized vector into cells, thus determining the site of plasmid insertion and often increasing the integration frequency. A set of transforming plasmids carrying the GFP coding sequence in three reading frames was used for transformation. The plasmids were constructed so that GFP could be expressed only under control of a host promoter. Living transformants expressing GFP spatially and temporally could be rapidly identified in a very large background of non-expressing cells and fruiting bodies. The phenotypes of representative mutants range from cells that cannot aggregate and initiate cell-cell interactions, through mutant fruiting bodies, to apparently wild-type fruiting bodies expressing GFP in all or a subpopulation of cells. The ability to screen mutant living cells and tissues for GFP expression is rapid and effective and likely to have application in many transformable systems where screening by gene and promoter trapping is essential for understanding temporal and spatial gene regulation.
Authors:
P Fey; E C Cox
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Current biology : CB     Volume:  7     ISSN:  0960-9822     ISO Abbreviation:  Curr. Biol.     Publication Date:  1997 Nov 
Date Detail:
Created Date:  1998-02-25     Completed Date:  1998-02-25     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9107782     Medline TA:  Curr Biol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  909-12     Citation Subset:  IM    
Affiliation:
Department of Molecular Biology, Princeton University, New Jersey 08544, USA.
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MeSH Terms
Descriptor/Qualifier:
Cloning, Molecular / methods*
Green Fluorescent Proteins
Luminescent Proteins / biosynthesis,  genetics*
Mutation*
Myxomycetes / genetics*,  growth & development*
Phenotype
Plasmids
Restriction Mapping
Transformation, Genetic
Chemical
Reg. No./Substance:
0/Luminescent Proteins; 147336-22-9/Green Fluorescent Proteins

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


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