Production and Purification of a Recombinant Elastomeric Polypeptide, G-(VPGVG)19-VPGV, from Escherichia coli
An elastomeric polypeptide was produced, with the sequence G‐(VPGVG)19‐VPGV, as a fusion to glutathione S‐transferase using the vector pGEX‐3X. The fusion protein was expressed to high levels in Escherichia coli as indicated by SDS‐PAGE analysis of induced cells. The fusion protein was affinity puri...
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| Veröffentlicht in: | Biotechnology progress 1992-07, Vol.8 (4), p.347-352 |
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| creator | Mcpherson, David T. Morrow, Casey Minehan, Daniel S. Wu, Jianguo Hunter, Eric Urry, Dan W. |
| description | An elastomeric polypeptide was produced, with the sequence G‐(VPGVG)19‐VPGV, as a fusion to glutathione S‐transferase using the vector pGEX‐3X. The fusion protein was expressed to high levels in Escherichia coli as indicated by SDS‐PAGE analysis of induced cells. The fusion protein was affinity purified and cleaved with protease factor Xa, and the elastomeric polypeptide was recovered to a high degree of purity as indicated by SDS—PAGE followed by staining with CuCl2. The physical characterizations of carbon‐13 and proton nuclear magnetic resonance and of the temperature profile for turbidity formation for the inverse temperature transition of hydrophobic folding and assembly attest to the successful microbial synthesis of the polypentapeptide of elastin. The results of these studies provide the initial progress toward achieving a more economical and practical means of producing material for elastic protein‐based polymer research and applications. |
| doi_str_mv | 10.1021/bp00016a012 |
| format | Article |
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The fusion protein was expressed to high levels in Escherichia coli as indicated by SDS‐PAGE analysis of induced cells. The fusion protein was affinity purified and cleaved with protease factor Xa, and the elastomeric polypeptide was recovered to a high degree of purity as indicated by SDS—PAGE followed by staining with CuCl2. The physical characterizations of carbon‐13 and proton nuclear magnetic resonance and of the temperature profile for turbidity formation for the inverse temperature transition of hydrophobic folding and assembly attest to the successful microbial synthesis of the polypentapeptide of elastin. The results of these studies provide the initial progress toward achieving a more economical and practical means of producing material for elastic protein‐based polymer research and applications.</description><identifier>ISSN: 8756-7938</identifier><identifier>EISSN: 1520-6033</identifier><identifier>DOI: 10.1021/bp00016a012</identifier><identifier>PMID: 1368456</identifier><identifier>CODEN: BIPRET</identifier><language>eng</language><publisher>USA: American Chemical Society</publisher><subject>Amino Acid Sequence ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Cloning, Molecular ; DNA, Bacterial ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli - metabolism ; Fundamental and applied biological sciences. Psychology ; Genes, Synthetic ; Genetic engineering ; Genetic technics ; Magnetic Resonance Spectroscopy ; Methods. Procedures. Technologies ; Modification of gene expression level ; Molecular Sequence Data ; Peptide Biosynthesis ; Peptides - isolation & purification ; Polymerase Chain Reaction ; Recombinant Fusion Proteins - biosynthesis ; Recombinant Fusion Proteins - isolation & purification</subject><ispartof>Biotechnology progress, 1992-07, Vol.8 (4), p.347-352</ispartof><rights>Copyright © 1992 American Institute of Chemical Engineers (AIChE)</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5012-d6dda106b8826f6cbfe7cac8fe164a81eb598030e18916dcdf963ad288fed2873</citedby><cites>FETCH-LOGICAL-c5012-d6dda106b8826f6cbfe7cac8fe164a81eb598030e18916dcdf963ad288fed2873</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,2764,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4360682$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1368456$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mcpherson, David T.</creatorcontrib><creatorcontrib>Morrow, Casey</creatorcontrib><creatorcontrib>Minehan, Daniel S.</creatorcontrib><creatorcontrib>Wu, Jianguo</creatorcontrib><creatorcontrib>Hunter, Eric</creatorcontrib><creatorcontrib>Urry, Dan W.</creatorcontrib><title>Production and Purification of a Recombinant Elastomeric Polypeptide, G-(VPGVG)19-VPGV, from Escherichia coli</title><title>Biotechnology progress</title><addtitle>Biotechnol Progress</addtitle><description>An elastomeric polypeptide was produced, with the sequence G‐(VPGVG)19‐VPGV, as a fusion to glutathione S‐transferase using the vector pGEX‐3X. The fusion protein was expressed to high levels in Escherichia coli as indicated by SDS‐PAGE analysis of induced cells. The fusion protein was affinity purified and cleaved with protease factor Xa, and the elastomeric polypeptide was recovered to a high degree of purity as indicated by SDS—PAGE followed by staining with CuCl2. The physical characterizations of carbon‐13 and proton nuclear magnetic resonance and of the temperature profile for turbidity formation for the inverse temperature transition of hydrophobic folding and assembly attest to the successful microbial synthesis of the polypentapeptide of elastin. The results of these studies provide the initial progress toward achieving a more economical and practical means of producing material for elastic protein‐based polymer research and applications.</description><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cloning, Molecular</subject><subject>DNA, Bacterial</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Escherichia coli - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Synthetic</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Magnetic Resonance Spectroscopy</subject><subject>Methods. Procedures. Technologies</subject><subject>Modification of gene expression level</subject><subject>Molecular Sequence Data</subject><subject>Peptide Biosynthesis</subject><subject>Peptides - isolation & purification</subject><subject>Polymerase Chain Reaction</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Recombinant Fusion Proteins - isolation & purification</subject><issn>8756-7938</issn><issn>1520-6033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1v1DAQhi1EVZbCiTOSDwiBaMCO448c22pJkaoSrUqRuFiOP1RDEgc7Eey_J2lWhROX8WjmmXfs1wC8wOg9Rjn-0AwIIcwUwvkjsME0RxlDhDwGG8Epy3hJxBPwNKXvMyYQy4_BMSZMFJRtQFfHYCY9-tBD1RtYT9E7r9V9ITio4M7q0DW-V_0It61KY-hs9BrWod0Pdhi9saewyt7c1tVt9RaX2ZKcQhdDB7dJ3y3wnVdQh9Y_A0dOtck-P5wn4MvH7c3FZXb1ufp0cXaVaTo_IjPMGIURa4TImWO6cZZrpYWzmBVKYNvQUiCCLBYlZkYbVzKiTC5mYo6cnIDXq-4Qw8_JplF2Pmnbtqq3YUqSE0xnJ-gMvltBHUNK0To5RN-puJcYycVc-Y-5M_3yIDs1nTV_2dXNuf_q0FdJq9ZF1WufHrCCMMTEIsNX7Jdv7f5_G-X5Tb2jRb78W3F_gWyd9Gm0vx8mVfwhGSecyq_XldwVlJffyLVE5A-Chp_S</recordid><startdate>199207</startdate><enddate>199207</enddate><creator>Mcpherson, David T.</creator><creator>Morrow, Casey</creator><creator>Minehan, Daniel S.</creator><creator>Wu, Jianguo</creator><creator>Hunter, Eric</creator><creator>Urry, Dan W.</creator><general>American Chemical Society</general><general>American Institute of Chemical Engineers</general><scope>BSCLL</scope><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>7X8</scope></search><sort><creationdate>199207</creationdate><title>Production and Purification of a Recombinant Elastomeric Polypeptide, G-(VPGVG)19-VPGV, from Escherichia coli</title><author>Mcpherson, David T. ; Morrow, Casey ; Minehan, Daniel S. ; Wu, Jianguo ; Hunter, Eric ; Urry, Dan W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5012-d6dda106b8826f6cbfe7cac8fe164a81eb598030e18916dcdf963ad288fed2873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cloning, Molecular</topic><topic>DNA, Bacterial</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Escherichia coli - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Synthetic</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Magnetic Resonance Spectroscopy</topic><topic>Methods. Procedures. Technologies</topic><topic>Modification of gene expression level</topic><topic>Molecular Sequence Data</topic><topic>Peptide Biosynthesis</topic><topic>Peptides - isolation & purification</topic><topic>Polymerase Chain Reaction</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Recombinant Fusion Proteins - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mcpherson, David T.</creatorcontrib><creatorcontrib>Morrow, Casey</creatorcontrib><creatorcontrib>Minehan, Daniel S.</creatorcontrib><creatorcontrib>Wu, Jianguo</creatorcontrib><creatorcontrib>Hunter, Eric</creatorcontrib><creatorcontrib>Urry, Dan W.</creatorcontrib><collection>Istex</collection><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>MEDLINE - Academic</collection><jtitle>Biotechnology progress</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mcpherson, David T.</au><au>Morrow, Casey</au><au>Minehan, Daniel S.</au><au>Wu, Jianguo</au><au>Hunter, Eric</au><au>Urry, Dan W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production and Purification of a Recombinant Elastomeric Polypeptide, G-(VPGVG)19-VPGV, from Escherichia coli</atitle><jtitle>Biotechnology progress</jtitle><addtitle>Biotechnol Progress</addtitle><date>1992-07</date><risdate>1992</risdate><volume>8</volume><issue>4</issue><spage>347</spage><epage>352</epage><pages>347-352</pages><issn>8756-7938</issn><eissn>1520-6033</eissn><coden>BIPRET</coden><abstract>An elastomeric polypeptide was produced, with the sequence G‐(VPGVG)19‐VPGV, as a fusion to glutathione S‐transferase using the vector pGEX‐3X. The fusion protein was expressed to high levels in Escherichia coli as indicated by SDS‐PAGE analysis of induced cells. The fusion protein was affinity purified and cleaved with protease factor Xa, and the elastomeric polypeptide was recovered to a high degree of purity as indicated by SDS—PAGE followed by staining with CuCl2. The physical characterizations of carbon‐13 and proton nuclear magnetic resonance and of the temperature profile for turbidity formation for the inverse temperature transition of hydrophobic folding and assembly attest to the successful microbial synthesis of the polypentapeptide of elastin. The results of these studies provide the initial progress toward achieving a more economical and practical means of producing material for elastic protein‐based polymer research and applications.</abstract><cop>USA</cop><pub>American Chemical Society</pub><pmid>1368456</pmid><doi>10.1021/bp00016a012</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 8756-7938 |
| ispartof | Biotechnology progress, 1992-07, Vol.8 (4), p.347-352 |
| issn | 8756-7938 1520-6033 |
| language | eng |
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| source | MEDLINE; ACS Publications |
| subjects | Amino Acid Sequence Base Sequence Biological and medical sciences Biotechnology Cloning, Molecular DNA, Bacterial Electrophoresis, Polyacrylamide Gel Escherichia coli - metabolism Fundamental and applied biological sciences. Psychology Genes, Synthetic Genetic engineering Genetic technics Magnetic Resonance Spectroscopy Methods. Procedures. Technologies Modification of gene expression level Molecular Sequence Data Peptide Biosynthesis Peptides - isolation & purification Polymerase Chain Reaction Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - isolation & purification |
| title | Production and Purification of a Recombinant Elastomeric Polypeptide, G-(VPGVG)19-VPGV, from Escherichia coli |
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