pLEF, a novel vector for expression of glutathione S-transferase fusion proteins in mammalian cells
An expression vector, pLEF, has been used to produce the intracellular domain (IC) of the human CD95 (Fas/APO-1) apoptosis receptor as a glutathione S-transferase (GST) fusion protein in murine L929 fibroblasts. GST::CD95IC was affinity-purified in a single step using glutathione-Sepharose. Purifica...
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| Veröffentlicht in: | Gene 1996-03, Vol.169 (2), p.281-282 |
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| container_end_page | 282 |
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| container_issue | 2 |
| container_start_page | 281 |
| container_title | Gene |
| container_volume | 169 |
| creator | Rudert, Fritz Visser, Elizabeth Gradl, Gabriele Grandison, Prudence Shemshedini, Lirim Wang, Yue Grierson, Alastair Watson, James |
| description | An expression vector, pLEF, has been used to produce the intracellular domain (IC) of the human CD95 (Fas/APO-1) apoptosis receptor as a glutathione S-transferase (GST) fusion protein in murine L929 fibroblasts. GST::CD95IC was affinity-purified in a single step using glutathione-Sepharose. Purification of GST::CD95IC from
32P-labelled L929 cells and cleavage with thrombin revealed that CD95IC was phosphorylated in vivo when produced as a GST fusion protein. Therefore, pLEF may facilitate the mapping of in vivo-modified sites of eukaryotic proteins |
| doi_str_mv | 10.1016/0378-1119(95)00820-9 |
| format | Article |
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32P-labelled L929 cells and cleavage with thrombin revealed that CD95IC was phosphorylated in vivo when produced as a GST fusion protein. Therefore, pLEF may facilitate the mapping of in vivo-modified sites of eukaryotic proteins</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>apoptosis receptor</subject><subject>Base Sequence</subject><subject>Cell Line</subject><subject>DNA</subject><subject>fas Receptor - chemistry</subject><subject>fas Receptor - genetics</subject><subject>fas Receptor - isolation & purification</subject><subject>Gene Expression - genetics</subject><subject>Genetic Vectors - genetics</subject><subject>Glutathione Transferase - genetics</subject><subject>Humans</subject><subject>Mammals</subject><subject>Mice</subject><subject>Molecular Sequence Data</subject><subject>neomycin selection</subject><subject>phosphorylation</subject><subject>Protein purification</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - isolation & purification</subject><subject>Thrombin - metabolism</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkVtLBSEUhSWKOl3-QYFPUdCUzuioL0FENzjQQ_UsjrMtYy4nnTnUv8_pHHosYSuyP5ebtRA6pOScElpekELIjFKqThQ_JUTmJFMbaEalUBkhhdxEs19kB-3G-E7S4jzfRtuyZIKV-QzZxfzm9gwb3PVLaPAS7NAH7FLB5yJAjL7vcO_wazMOZnhLN8BP2RBMFx0EEwG78YdZhH4A30XsO9yatjWNNx220DRxH20500Q4WJ976OX25vn6Pps_3j1cX80zy3I2ZK4ipTO1cCCZIJw6zqBMu63KnNWFFDknorZSWmMYTVWTvHCMQ1XldaKKPXS80k2zfIwQB936OE1gOujHqIVQSolC_QtSQajgBU8gW4E29DEGcHoRfGvCl6ZETyHoyWE9OawV1z8h6En_aK0_Vi3Uv4_Wrqf-5aoPyY2lh6Cj9dBZqH1IAei6939_8A077Zc7</recordid><startdate>19960309</startdate><enddate>19960309</enddate><creator>Rudert, Fritz</creator><creator>Visser, Elizabeth</creator><creator>Gradl, Gabriele</creator><creator>Grandison, Prudence</creator><creator>Shemshedini, Lirim</creator><creator>Wang, Yue</creator><creator>Grierson, Alastair</creator><creator>Watson, James</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19960309</creationdate><title>pLEF, a novel vector for expression of glutathione S-transferase fusion proteins in mammalian cells</title><author>Rudert, Fritz ; 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GST::CD95IC was affinity-purified in a single step using glutathione-Sepharose. Purification of GST::CD95IC from
32P-labelled L929 cells and cleavage with thrombin revealed that CD95IC was phosphorylated in vivo when produced as a GST fusion protein. Therefore, pLEF may facilitate the mapping of in vivo-modified sites of eukaryotic proteins</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>8647462</pmid><doi>10.1016/0378-1119(95)00820-9</doi><tpages>2</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1119 |
| ispartof | Gene, 1996-03, Vol.169 (2), p.281-282 |
| issn | 0378-1119 1879-0038 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Amino Acid Sequence Animals apoptosis receptor Base Sequence Cell Line DNA fas Receptor - chemistry fas Receptor - genetics fas Receptor - isolation & purification Gene Expression - genetics Genetic Vectors - genetics Glutathione Transferase - genetics Humans Mammals Mice Molecular Sequence Data neomycin selection phosphorylation Protein purification Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - isolation & purification Thrombin - metabolism |
| title | pLEF, a novel vector for expression of glutathione S-transferase fusion proteins in mammalian cells |
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