| RRC ID |
61223
|
| 著者 |
Miyoshi S, Motohashi T, Nakamura Y, Osawa M, Hiroyama T, Kim DK, Tokumoto Y, Nakauchi H.
|
| タイトル |
A transmembrane trap method for efficient cloning of genes encoding proteins possessing transmembrane domain.
|
| ジャーナル |
Biochem Biophys Res Commun
|
| Abstract |
To facilitate searching for genes encoding cell membrane proteins, we developed a method for isolating cDNAs that contain sequences for hydrophobic transmembrane runs. This cloning strategy, termed the "transmembrane (TM) trap method," utilizes a vector that directs the cell surface expression of mouse CD4 fusion protein when an insert encoding hydrophobic transmembrane sequences is cloned in-frame with correct orientation. We applied this novel method to isolation of cytokine receptor cDNAs. Our strategy enabled efficient isolation of relatively rare species encoding receptors such as IL-2Rgamma, IL-3Rbeta, IL-4Ralpha, IL-5Ralpha, and IL-6Ralpha. This method also could be used to isolate cDNAs for intracellular molecules with a transmembrane region, e.g., bcl-2. These results indicate that the TM trap method provides an efficient cloning strategy for identification of various families of genes encoding proteins with one or more transmembrane regions.
|
| 巻・号 |
289(5)
|
| ページ |
1192-8
|
| 公開日 |
2001-12-21
|
| DOI |
10.1006/bbrc.2001.6097
|
| PII |
S0006-291X(01)96097-8
|
| PMID |
11741319
|
| MeSH |
Animals
Base Sequence
CD4 Antigens / genetics
COS Cells
Cell Line
Cell Membrane / chemistry
Cloning, Molecular / methods*
DNA, Complementary / genetics
DNA, Complementary / isolation & purification
Gene Expression
Gene Library
Genetic Vectors
Humans
Membrane Proteins / chemistry*
Membrane Proteins / genetics*
Mice
Plasmids / genetics
Protein Structure, Tertiary
Recombinant Fusion Proteins / genetics
Transfection
|
| IF |
2.985
|
| リソース情報 |
| ヒト・動物細胞 |
F-36P(RCB0775) |
