Reference - Detail
| RRC ID | 30952 |
|---|---|
| Author | Levi S, Polyakov M, Egelhoff TT. |
| Title | Green fluorescent protein and epitope tag fusion vectors for Dictyostelium discoideum. |
| Journal | Plasmid |
| Abstract |
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. |
| Volume | 44(3) |
| Pages | 231-8 |
| Published | 2000-11-1 |
| DOI | 10.1006/plas.2000.1487 |
| PII | S0147-619X(00)91487-2 |
| PMID | 11078649 |
| MeSH | Amino Acid Sequence Animals Bacterial Proteins / genetics* Bacterial Proteins / metabolism Base Sequence Cloning, Molecular / methods* Dictyostelium / genetics* Dictyostelium / metabolism Escherichia coli Proteins* Genetic Vectors / genetics* Green Fluorescent Proteins Luminescent Proteins / biosynthesis Luminescent Proteins / genetics* Molecular Sequence Data Oligopeptides Oxidoreductases* Peptides / genetics* Peptides / metabolism Plasmids / genetics* Recombinant Fusion Proteins / biosynthesis Recombinant Fusion Proteins / genetics* |
| IF | 2.805 |
| Times Cited | 130 |
| WOS Category | MICROBIOLOGY GENETICS & HEREDITY |
| Resource | |
| Cellular slime molds | G90010 |