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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
3
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pubmed:dateCreated |
2000-12-13
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pubmed:databankReference |
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pubmed:abstractText |
We have constructed expression vectors for Dictyostelium discoideum which encode a green fluorescent protein (GFP) sequence upstream of a multicloning site for introduction of sequences of interest. Insertion of cDNAs into the multicloning site results in expression of fusion protein bearing an amino- or carboxyl-terminal GFP tag which can be used for fluorescent localization studies in Dictyostelium cells. A parallel construct fuses a FLAG epitope tag at the amino terminus of expressed protein. Each fusion cartridge was placed either in a G418-resistance vector allowing transactivated Ddp2-based extrachromosomal replication or in a vector allowing autonomous Ddp1-based replication. Distinct differences in expression stability were observed in the two vector types. When GFP-expressing cells were analyzed by fluorescence microscopy, significant cell-to-cell variability in expression level was observed when expression was based on the Ddp2 vector, while less cell-to-cell variation in expression level was observed when the Ddp1 backbone was used for expression.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0147-619X
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pubmed:author |
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pubmed:copyrightInfo |
Copyright 2000 Academic Press.
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pubmed:issnType |
Print
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pubmed:volume |
44
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
231-8
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:11078649-Amino Acid Sequence,
pubmed-meshheading:11078649-Animals,
pubmed-meshheading:11078649-Bacterial Proteins,
pubmed-meshheading:11078649-Base Sequence,
pubmed-meshheading:11078649-Cloning, Molecular,
pubmed-meshheading:11078649-Dictyostelium,
pubmed-meshheading:11078649-Escherichia coli Proteins,
pubmed-meshheading:11078649-Genetic Vectors,
pubmed-meshheading:11078649-Green Fluorescent Proteins,
pubmed-meshheading:11078649-Luminescent Proteins,
pubmed-meshheading:11078649-Molecular Sequence Data,
pubmed-meshheading:11078649-Oxidoreductases,
pubmed-meshheading:11078649-Peptides,
pubmed-meshheading:11078649-Plasmids,
pubmed-meshheading:11078649-Recombinant Fusion Proteins
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pubmed:year |
2000
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pubmed:articleTitle |
Green fluorescent protein and epitope tag fusion vectors for Dictyostelium discoideum.
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pubmed:affiliation |
Department of Physiology and Biophysics, Case Western Reserve School of Medicine, Cleveland, Ohio 44106-4970, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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