論文 - 詳細
RRC ID | 30952 |
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著者 | Levi S, Polyakov M, Egelhoff TT. |
タイトル | Green fluorescent protein and epitope tag fusion vectors for Dictyostelium discoideum. |
ジャーナル | 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. |
巻・号 | 44(3) |
ページ | 231-8 |
公開日 | 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 |
引用数 | 130 |
WOS 分野 | MICROBIOLOGY GENETICS & HEREDITY |
リソース情報 | |
細胞性粘菌 | G90010 |