Gene expression in a cell-free system on the preparative scale
A cell-free system for preparative gene expression is described. It is composed of DNA-free Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works...
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| Veröffentlicht in: | Gene 1989-12, Vol.84 (2), p.463-466 |
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| Format: | Artikel |
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| container_end_page | 466 |
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| container_issue | 2 |
| container_start_page | 463 |
| container_title | Gene |
| container_volume | 84 |
| creator | Baranov, Vladimir I. Morozov, Igor Yu Ortlepp, Stephen A. Spirin, Alexander S. |
| description | A cell-free system for preparative gene expression is described. It is composed of DNA-free
Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works at a high constant rate for tens of hours. The yield of synthesised proteins after 20–50 h is hundreds of μg from 1 ml of the reaction mixture. Electrophoretic analysis of translation products confirms synthesis of proteins of the expected molecular mass. |
| doi_str_mv | 10.1016/0378-1119(89)90521-0 |
| format | Article |
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Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works at a high constant rate for tens of hours. The yield of synthesised proteins after 20–50 h is hundreds of μg from 1 ml of the reaction mixture. Electrophoretic analysis of translation products confirms synthesis of proteins of the expected molecular mass.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>beta-Lactamases - biosynthesis</subject><subject>beta-Lactamases - genetics</subject><subject>Biological and medical sciences</subject><subject>Buffers</subject><subject>continuous flow</subject><subject>Coupled transcription-translation</subject><subject>Culture Media</subject><subject>dihydrofolic acid reductase</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Guanosine Triphosphate - metabolism</subject><subject>in vitro system</subject><subject>Kinetics</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Nucleotides</subject><subject>Plasmids</subject><subject>Protein Biosynthesis</subject><subject>Tetrahydrofolate Dehydrogenase - genetics</subject><subject>Transcription, Genetic</subject><subject>β-lactamase</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkF1LwzAUhoMoc07_gUIvRPSiepK0TXMzkOEXDLzR65Ampxjpx0y64f69rSu71NwE8j7nzeEh5JzCLQWa3QEXeUwplde5vJGQMhrDAZnSXMgYgOeHZLpHjslJCJ_QnzRlEzJhmeSMplMyf8IGI_xeeQzBtU3kmkhHBqsqLj1iFLahwzrqg-4Do55aaa87t-kToys8JUelrgKejfeMvD8-vC2e4-Xr08vifhmbJOVdXIrEFrT_UxdMC5ObUgPXrLDW9q-AnErkmkIpRaG5TSwwy4pccAM84xb4jFztele-_Vpj6FTtwrClbrBdByVkknBgyb8gTRORSTo0JjvQ-DYEj6VaeVdrv1UU1OBXDfLUIE_lUv36VcPYxdi_Lmq0-6FRaJ9fjrkeBJVeN8aFPZYJSGk-YPMdhr20jUOvgnHYGLTOo-mUbd3fe_wA8t2VPw</recordid><startdate>19891214</startdate><enddate>19891214</enddate><creator>Baranov, Vladimir I.</creator><creator>Morozov, Igor Yu</creator><creator>Ortlepp, Stephen A.</creator><creator>Spirin, Alexander S.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>19891214</creationdate><title>Gene expression in a cell-free system on the preparative scale</title><author>Baranov, Vladimir I. ; Morozov, Igor Yu ; Ortlepp, Stephen A. ; Spirin, Alexander S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c453t-f74db1693ab2a7c8cfa03a2bddd1690e319e3a10f97ba3d4d02d2b873c0363d03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>beta-Lactamases - biosynthesis</topic><topic>beta-Lactamases - genetics</topic><topic>Biological and medical sciences</topic><topic>Buffers</topic><topic>continuous flow</topic><topic>Coupled transcription-translation</topic><topic>Culture Media</topic><topic>dihydrofolic acid reductase</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Guanosine Triphosphate - metabolism</topic><topic>in vitro system</topic><topic>Kinetics</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Nucleotides</topic><topic>Plasmids</topic><topic>Protein Biosynthesis</topic><topic>Tetrahydrofolate Dehydrogenase - genetics</topic><topic>Transcription, Genetic</topic><topic>β-lactamase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Baranov, Vladimir I.</creatorcontrib><creatorcontrib>Morozov, Igor Yu</creatorcontrib><creatorcontrib>Ortlepp, Stephen A.</creatorcontrib><creatorcontrib>Spirin, Alexander S.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Baranov, Vladimir I.</au><au>Morozov, Igor Yu</au><au>Ortlepp, Stephen A.</au><au>Spirin, Alexander S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene expression in a cell-free system on the preparative scale</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1989-12-14</date><risdate>1989</risdate><volume>84</volume><issue>2</issue><spage>463</spage><epage>466</epage><pages>463-466</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>A cell-free system for preparative gene expression is described. 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Escherichia coli extract and added plasmid DNA; coupled transcription-translation proceeds with a continuous flow of the feeding solution containing nucleoside triphosphates and amino acids. The system works at a high constant rate for tens of hours. The yield of synthesised proteins after 20–50 h is hundreds of μg from 1 ml of the reaction mixture. Electrophoretic analysis of translation products confirms synthesis of proteins of the expected molecular mass.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>2693215</pmid><doi>10.1016/0378-1119(89)90521-0</doi><tpages>4</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1119 |
| ispartof | Gene, 1989-12, Vol.84 (2), p.463-466 |
| issn | 0378-1119 1879-0038 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Adenosine Triphosphate - metabolism beta-Lactamases - biosynthesis beta-Lactamases - genetics Biological and medical sciences Buffers continuous flow Coupled transcription-translation Culture Media dihydrofolic acid reductase Electrophoresis, Polyacrylamide Gel Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Expression Guanosine Triphosphate - metabolism in vitro system Kinetics Molecular and cellular biology Molecular genetics Nucleotides Plasmids Protein Biosynthesis Tetrahydrofolate Dehydrogenase - genetics Transcription, Genetic β-lactamase |
| title | Gene expression in a cell-free system on the preparative scale |
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