Characterization of the Foot-and-Mouth Disease Virus 3C Protease Expressed in Escherichia coli
We have constructed a clone encoding the foot-and-mouth disease virus (FMDV) 3C protease gene (p3C) using the polymerase chain reaction. The construct was engineered to contain initiation and termination codons and cloned into a plasmid under the control of the bacteriophage T7 promoter. The p3C gen...
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| Veröffentlicht in: | Virology (New York, N.Y.) N.Y.), 1993-11, Vol.197 (1), p.320-327 |
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| creator | Bablanian, Gayne M. Grubman, Marvin J. |
| description | We have constructed a clone encoding the foot-and-mouth disease virus (FMDV) 3C protease gene (p3C) using the polymerase chain reaction. The construct was engineered to contain initiation and termination codons and cloned into a plasmid under the control of the bacteriophage T7 promoter. The p3C gene was expressed both in an
in vitro transcription-translation system and
in vivo in an
Escherichia coli system containing an inducible T7 RNA polymerase gene. In both systems the expressed products were of the appropriate molecular weight and immunologically reactive with bovine convalescent serum.
E. coli -expressed 3C protein was mainly found in the insoluble fraction of cell lysates. The
E. coli-expressed protease was assayed in an
in vitro system with radiolabeled P1 capsid precursor protein and P2 precursor protein as substrates.
E. coli-expressed 3C completely processed the P1 and P2 precursors into mature capsid and nonstructural proteins, respectively. The kinetics of processing of P1 by
E. coli-expressed 3C revealed the following order of cleavage: VP3-VP1, VPO-VP3, VP1-2A. |
| doi_str_mv | 10.1006/viro.1993.1593 |
| format | Article |
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in vitro transcription-translation system and
in vivo in an
Escherichia coli system containing an inducible T7 RNA polymerase gene. In both systems the expressed products were of the appropriate molecular weight and immunologically reactive with bovine convalescent serum.
E. coli -expressed 3C protein was mainly found in the insoluble fraction of cell lysates. The
E. coli-expressed protease was assayed in an
in vitro system with radiolabeled P1 capsid precursor protein and P2 precursor protein as substrates.
E. coli-expressed 3C completely processed the P1 and P2 precursors into mature capsid and nonstructural proteins, respectively. The kinetics of processing of P1 by
E. coli-expressed 3C revealed the following order of cleavage: VP3-VP1, VPO-VP3, VP1-2A.</description><identifier>ISSN: 0042-6822</identifier><identifier>EISSN: 1096-0341</identifier><identifier>DOI: 10.1006/viro.1993.1593</identifier><identifier>PMID: 8212567</identifier><identifier>CODEN: VIRLAX</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>3C Viral Proteases ; Aphthovirus - enzymology ; Aphthovirus - genetics ; Base Sequence ; Biological and medical sciences ; Cloning, Molecular ; Cysteine Endopeptidases - biosynthesis ; Cysteine Endopeptidases - genetics ; Cysteine Endopeptidases - isolation & purification ; DNA Primers ; Escherichia coli ; Escherichia coli - enzymology ; Escherichia coli - genetics ; foot-and-mouth disease virus ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genetics ; Kinetics ; Methionine - metabolism ; Microbiology ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; Protein Biosynthesis ; Recombinant Proteins - metabolism ; Transcription, Genetic ; Viral Proteins ; Virology</subject><ispartof>Virology (New York, N.Y.), 1993-11, Vol.197 (1), p.320-327</ispartof><rights>1993 Academic Press</rights><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c399t-4eb12be0e74f8331898cab73879e17fbe87238983b55708c0d3ce21b20bfe98b3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1006/viro.1993.1593$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3839101$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8212567$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bablanian, Gayne M.</creatorcontrib><creatorcontrib>Grubman, Marvin J.</creatorcontrib><title>Characterization of the Foot-and-Mouth Disease Virus 3C Protease Expressed in Escherichia coli</title><title>Virology (New York, N.Y.)</title><addtitle>Virology</addtitle><description>We have constructed a clone encoding the foot-and-mouth disease virus (FMDV) 3C protease gene (p3C) using the polymerase chain reaction. The construct was engineered to contain initiation and termination codons and cloned into a plasmid under the control of the bacteriophage T7 promoter. The p3C gene was expressed both in an
in vitro transcription-translation system and
in vivo in an
Escherichia coli system containing an inducible T7 RNA polymerase gene. In both systems the expressed products were of the appropriate molecular weight and immunologically reactive with bovine convalescent serum.
E. coli -expressed 3C protein was mainly found in the insoluble fraction of cell lysates. The
E. coli-expressed protease was assayed in an
in vitro system with radiolabeled P1 capsid precursor protein and P2 precursor protein as substrates.
E. coli-expressed 3C completely processed the P1 and P2 precursors into mature capsid and nonstructural proteins, respectively. The kinetics of processing of P1 by
E. coli-expressed 3C revealed the following order of cleavage: VP3-VP1, VPO-VP3, VP1-2A.</description><subject>3C Viral Proteases</subject><subject>Aphthovirus - enzymology</subject><subject>Aphthovirus - genetics</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>Cysteine Endopeptidases - biosynthesis</subject><subject>Cysteine Endopeptidases - genetics</subject><subject>Cysteine Endopeptidases - isolation & purification</subject><subject>DNA Primers</subject><subject>Escherichia coli</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli - genetics</subject><subject>foot-and-mouth disease virus</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetics</subject><subject>Kinetics</subject><subject>Methionine - metabolism</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Plasmids</subject><subject>Protein Biosynthesis</subject><subject>Recombinant Proteins - metabolism</subject><subject>Transcription, Genetic</subject><subject>Viral Proteins</subject><subject>Virology</subject><issn>0042-6822</issn><issn>1096-0341</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEFv1DAQRi0EKtvClRuSD4hblrGdxPYRbbcFqVV7AI5YjjNRjLLxYjsV5dfjZVe9IU6jmXnzafQIecNgzQDaDw8-hjXTWqxZo8UzsmKg2wpEzZ6TFUDNq1Zx_pKcp_QDSi8lnJEzxRlvWrki3zejjdZljP63zT7MNAw0j0ivQsiVnfvqNix5pJc-oU1Iv_m4JCo29D6G_Hey_bWPmBL21M90m9xYotzoLXVh8q_Ii8FOCV-f6gX5erX9svlU3dxdf958vKmc0DpXNXaMdwgo60EJwZRWznZSKKmRyaFDJbkoQ9E1jQTloBcOOes4dANq1YkL8v6Yu4_h54Ipm51PDqfJzhiWZGQLvAbW_BdkbSEFQAHXR9DFkFLEweyj39n4aBiYg3lzMG8O5s3BfDl4e0peuh32T_hJddm_O-1tcnYaop2dT0-YUEIzYAVTRwyLrgeP0STncXbY-4gumz74f33wBx7gnpo</recordid><startdate>19931101</startdate><enddate>19931101</enddate><creator>Bablanian, Gayne M.</creator><creator>Grubman, Marvin J.</creator><general>Elsevier Inc</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>7QO</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19931101</creationdate><title>Characterization of the Foot-and-Mouth Disease Virus 3C Protease Expressed in Escherichia coli</title><author>Bablanian, Gayne M. ; Grubman, Marvin J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c399t-4eb12be0e74f8331898cab73879e17fbe87238983b55708c0d3ce21b20bfe98b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>3C Viral Proteases</topic><topic>Aphthovirus - enzymology</topic><topic>Aphthovirus - genetics</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>Cysteine Endopeptidases - biosynthesis</topic><topic>Cysteine Endopeptidases - genetics</topic><topic>Cysteine Endopeptidases - isolation & purification</topic><topic>DNA Primers</topic><topic>Escherichia coli</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli - genetics</topic><topic>foot-and-mouth disease virus</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetics</topic><topic>Kinetics</topic><topic>Methionine - metabolism</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Plasmids</topic><topic>Protein Biosynthesis</topic><topic>Recombinant Proteins - metabolism</topic><topic>Transcription, Genetic</topic><topic>Viral Proteins</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bablanian, Gayne M.</creatorcontrib><creatorcontrib>Grubman, Marvin J.</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>Biotechnology Research Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Virology (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bablanian, Gayne M.</au><au>Grubman, Marvin J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of the Foot-and-Mouth Disease Virus 3C Protease Expressed in Escherichia coli</atitle><jtitle>Virology (New York, N.Y.)</jtitle><addtitle>Virology</addtitle><date>1993-11-01</date><risdate>1993</risdate><volume>197</volume><issue>1</issue><spage>320</spage><epage>327</epage><pages>320-327</pages><issn>0042-6822</issn><eissn>1096-0341</eissn><coden>VIRLAX</coden><abstract>We have constructed a clone encoding the foot-and-mouth disease virus (FMDV) 3C protease gene (p3C) using the polymerase chain reaction. The construct was engineered to contain initiation and termination codons and cloned into a plasmid under the control of the bacteriophage T7 promoter. The p3C gene was expressed both in an
in vitro transcription-translation system and
in vivo in an
Escherichia coli system containing an inducible T7 RNA polymerase gene. In both systems the expressed products were of the appropriate molecular weight and immunologically reactive with bovine convalescent serum.
E. coli -expressed 3C protein was mainly found in the insoluble fraction of cell lysates. The
E. coli-expressed protease was assayed in an
in vitro system with radiolabeled P1 capsid precursor protein and P2 precursor protein as substrates.
E. coli-expressed 3C completely processed the P1 and P2 precursors into mature capsid and nonstructural proteins, respectively. The kinetics of processing of P1 by
E. coli-expressed 3C revealed the following order of cleavage: VP3-VP1, VPO-VP3, VP1-2A.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>8212567</pmid><doi>10.1006/viro.1993.1593</doi><tpages>8</tpages></addata></record> |
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| issn | 0042-6822 1096-0341 |
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| recordid | cdi_proquest_miscellaneous_76024015 |
| source | MEDLINE; ScienceDirect Journals (5 years ago - present); EZB-FREE-00999 freely available EZB journals |
| subjects | 3C Viral Proteases Aphthovirus - enzymology Aphthovirus - genetics Base Sequence Biological and medical sciences Cloning, Molecular Cysteine Endopeptidases - biosynthesis Cysteine Endopeptidases - genetics Cysteine Endopeptidases - isolation & purification DNA Primers Escherichia coli Escherichia coli - enzymology Escherichia coli - genetics foot-and-mouth disease virus Fundamental and applied biological sciences. Psychology Gene Expression Genetics Kinetics Methionine - metabolism Microbiology Molecular Sequence Data Molecular Weight Plasmids Protein Biosynthesis Recombinant Proteins - metabolism Transcription, Genetic Viral Proteins Virology |
| title | Characterization of the Foot-and-Mouth Disease Virus 3C Protease Expressed in Escherichia coli |
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