Position-independent expression of a human nerve growth factor-luciferase reporter gene cloned on a yeast artificial chromosome vector
Identifieur interne : 000296 ( Main/Corpus ); précédent : 000295; suivant : 000297Position-independent expression of a human nerve growth factor-luciferase reporter gene cloned on a yeast artificial chromosome vector
Auteurs : Fred A. M. Asselbergs ; Rita Grossenbacher ; Rainer Ortmann ; Bastian Hengerer ; Gary K. Mcmaster ; Esther Sutter ; Roland Widmer ; Frank BuxtonSource :
- Nucleic Acids Research [ 0305-1048 ] ; 1998-04.
Abstract
Two yeast artificial chromosomes containing the entire human nerve growth factor gene were isolated and mapped. By homologous recombination a luciferase gene was precisely engineered into the coding portion of the NGF gene and a neomycin selection marker was placed adjacent to one of the YAC telomeres. Expression of the YAC-based NGF reporter gene and a plasmid-based NGF reporter gene were compared with the regulation of endogenous mouse NGF protein in mouse L929 fibroblasts. In contrast to the plasmid-based reporter gene, expression and regulation of the YAC-based reporter gene was independent of the site of integration of the transgene. Basic fibroblast growth factor and okadaic acid stimulated expression of the YAC transgene, whereas transforming growth factor-β and dexamethasone inhibited it. Although cyclic AMP strongly stimulated production of the endogenous mouse NGF, no effect was seen on the human NGF reporter genes. Downregulation of the secretion of endogenous mouse NGF already occurred at an EC50 of 1–2 nM dexamethasone, but downregulation of the expression of NGF reporter genes occurred only at EC50 of 10 nM. This higher concentration was also required for upregulation of luciferase genes driven by the dexamethasone-inducible promoter of the mouse mammary tumor virus in L929 fibroblasts.
Url:
DOI: 10.1093/nar/26.7.1826
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Asselbergs</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: fred_am.asselbergs@pharma.novartis.com</mods:affiliation></affiliation></author><author><name sortKey="Grossenbacher, Rita" sort="Grossenbacher, Rita" uniqKey="Grossenbacher R" first="Rita" last="Grossenbacher">Rita Grossenbacher</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author><author><name sortKey="Ortmann, Rainer" sort="Ortmann, Rainer" uniqKey="Ortmann R" first="Rainer" last="Ortmann">Rainer Ortmann</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author><author><name sortKey="Hengerer, Bastian" sort="Hengerer, Bastian" uniqKey="Hengerer B" first="Bastian" last="Hengerer">Bastian Hengerer</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author><author><name sortKey="Mcmaster, Gary K" sort="Mcmaster, Gary K" uniqKey="Mcmaster G" first="Gary K." last="Mcmaster">Gary K. 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Mcmaster</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author><author><name sortKey="Sutter, Esther" sort="Sutter, Esther" uniqKey="Sutter E" first="Esther" last="Sutter">Esther Sutter</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author><author><name sortKey="Widmer, Roland" sort="Widmer, Roland" uniqKey="Widmer R" first="Roland" last="Widmer">Roland Widmer</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author><author><name sortKey="Buxton, Frank" sort="Buxton, Frank" uniqKey="Buxton F" first="Frank" last="Buxton">Frank Buxton</name><affiliation><mods:affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Nucleic Acids Research</title><title level="j" type="abbrev">Nucleic Acids Research</title><idno type="ISSN">0305-1048</idno><idno type="eISSN">1362-4962</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="1998-04">1998-04</date><biblScope unit="volume">26</biblScope><biblScope unit="issue">7</biblScope><biblScope unit="page" from="1826">1826</biblScope><biblScope unit="page" to="1833">1833</biblScope></imprint><idno type="ISSN">0305-1048</idno></series><idno type="istex">454B71EBFD140159884DA71C097860998154F95D</idno><idno type="DOI">10.1093/nar/26.7.1826</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0305-1048</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract">Two yeast artificial chromosomes containing the entire human nerve growth factor gene were isolated and mapped. By homologous recombination a luciferase gene was precisely engineered into the coding portion of the NGF gene and a neomycin selection marker was placed adjacent to one of the YAC telomeres. Expression of the YAC-based NGF reporter gene and a plasmid-based NGF reporter gene were compared with the regulation of endogenous mouse NGF protein in mouse L929 fibroblasts. In contrast to the plasmid-based reporter gene, expression and regulation of the YAC-based reporter gene was independent of the site of integration of the transgene. Basic fibroblast growth factor and okadaic acid stimulated expression of the YAC transgene, whereas transforming growth factor-β and dexamethasone inhibited it. Although cyclic AMP strongly stimulated production of the endogenous mouse NGF, no effect was seen on the human NGF reporter genes. Downregulation of the secretion of endogenous mouse NGF already occurred at an EC50 of 1–2 nM dexamethasone, but downregulation of the expression of NGF reporter genes occurred only at EC50 of 10 nM. This higher concentration was also required for upregulation of luciferase genes driven by the dexamethasone-inducible promoter of the mouse mammary tumor virus in L929 fibroblasts.</div></front></TEI><istex><corpusName>oup</corpusName><author><json:item><name>Fred A.M. Asselbergs</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string><json:string>E-mail: fred_am.asselbergs@pharma.novartis.com</json:string></affiliations></json:item><json:item><name>Rita Grossenbacher</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item><json:item><name>Rainer Ortmann</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item><json:item><name>Bastian Hengerer</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item><json:item><name>Gary K. McMaster</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item><json:item><name>Esther Sutter</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item><json:item><name>Roland Widmer</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item><json:item><name>Frank Buxton</name><affiliations><json:string>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Articles</value></json:item></subject><language><json:string>eng</json:string></language><abstract>Two yeast artificial chromosomes containing the entire human nerve growth factor gene were isolated and mapped. By homologous recombination a luciferase gene was precisely engineered into the coding portion of the NGF gene and a neomycin selection marker was placed adjacent to one of the YAC telomeres. Expression of the YAC-based NGF reporter gene and a plasmid-based NGF reporter gene were compared with the regulation of endogenous mouse NGF protein in mouse L929 fibroblasts. In contrast to the plasmid-based reporter gene, expression and regulation of the YAC-based reporter gene was independent of the site of integration of the transgene. Basic fibroblast growth factor and okadaic acid stimulated expression of the YAC transgene, whereas transforming growth factor-β and dexamethasone inhibited it. Although cyclic AMP strongly stimulated production of the endogenous mouse NGF, no effect was seen on the human NGF reporter genes. Downregulation of the secretion of endogenous mouse NGF already occurred at an EC50 of 1–2 nM dexamethasone, but downregulation of the expression of NGF reporter genes occurred only at EC50 of 10 nM. This higher concentration was also required for upregulation of luciferase genes driven by the dexamethasone-inducible promoter of the mouse mammary tumor virus in L929 fibroblasts.</abstract><qualityIndicators><score>7.388</score><pdfVersion>1.2</pdfVersion><pdfPageSize>595 x 842 pts (A4)</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>1</keywordCount><abstractCharCount>1321</abstractCharCount><pdfWordCount>5769</pdfWordCount><pdfCharCount>36941</pdfCharCount><pdfPageCount>8</pdfPageCount><abstractWordCount>199</abstractWordCount></qualityIndicators><title>Position-independent expression of a human nerve growth factor-luciferase reporter gene cloned on a yeast artificial chromosome 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type="first">Rainer</forename><surname>Ortmann</surname></persName><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation></author><author><persName><forename type="first">Bastian</forename><surname>Hengerer</surname></persName><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation></author><author><persName><forename type="first">Gary K.</forename><surname>McMaster</surname></persName><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation></author><author><persName><forename type="first">Esther</forename><surname>Sutter</surname></persName><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation></author><author><persName><forename type="first">Roland</forename><surname>Widmer</surname></persName><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation></author><author><persName><forename type="first">Frank</forename><surname>Buxton</surname></persName><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation></author></analytic><monogr><title level="j">Nucleic Acids Research</title><title level="j" type="abbrev">Nucleic Acids Research</title><idno type="JournalID">nar</idno><idno type="pISSN">0305-1048</idno><idno type="eISSN">1362-4962</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="1998-04"></date><biblScope unit="volume">26</biblScope><biblScope unit="issue">7</biblScope><biblScope unit="page" from="1826">1826</biblScope><biblScope unit="page" to="1833">1833</biblScope></imprint></monogr><idno type="istex">454B71EBFD140159884DA71C097860998154F95D</idno><idno type="DOI">10.1093/nar/26.7.1826</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1998</date></creation><langUsage><language ident="en">en</language></langUsage><abstract><p>Two yeast artificial chromosomes containing the entire human nerve growth factor gene were isolated and mapped. By homologous recombination a luciferase gene was precisely engineered into the coding portion of the NGF gene and a neomycin selection marker was placed adjacent to one of the YAC telomeres. Expression of the YAC-based NGF reporter gene and a plasmid-based NGF reporter gene were compared with the regulation of endogenous mouse NGF protein in mouse L929 fibroblasts. In contrast to the plasmid-based reporter gene, expression and regulation of the YAC-based reporter gene was independent of the site of integration of the transgene. Basic fibroblast growth factor and okadaic acid stimulated expression of the YAC transgene, whereas transforming growth factor-β and dexamethasone inhibited it. Although cyclic AMP strongly stimulated production of the endogenous mouse NGF, no effect was seen on the human NGF reporter genes. Downregulation of the secretion of endogenous mouse NGF already occurred at an EC50 of 1–2 nM dexamethasone, but downregulation of the expression of NGF reporter genes occurred only at EC50 of 10 nM. This higher concentration was also required for upregulation of luciferase genes driven by the dexamethasone-inducible promoter of the mouse mammary tumor virus in L929 fibroblasts.</p></abstract><textClass><keywords scheme="keyword"><list><head>heading</head><item><term>Articles</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1998-04">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/454B71EBFD140159884DA71C097860998154F95D/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus oup" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article article-type="research-article"><front><journal-meta><journal-id journal-id-type="hwp">nar</journal-id><journal-id journal-id-type="publisher-id">nar</journal-id><journal-title>Nucleic Acids Research</journal-title><abbrev-journal-title>Nucleic Acids Research</abbrev-journal-title><issn pub-type="ppub">0305-1048</issn><issn pub-type="epub">1362-4962</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.1093/nar/26.7.1826</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group></article-categories><title-group><article-title>Position-independent expression of a human nerve growth factor-luciferase reporter gene cloned on a yeast artificial chromosome vector</article-title></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name><surname>Asselbergs</surname><given-names>Fred A.M.</given-names></name><xref ref-type="corresp" rid="cor1">*</xref></contrib><contrib contrib-type="author"><name><surname>Grossenbacher</surname><given-names>Rita</given-names></name></contrib><contrib contrib-type="author"><name><surname>Ortmann</surname><given-names>Rainer</given-names></name></contrib><contrib contrib-type="author"><name><surname>Hengerer</surname><given-names>Bastian</given-names></name></contrib><contrib contrib-type="author"><name><surname>McMaster</surname><given-names>Gary K.</given-names></name></contrib><contrib contrib-type="author"><name><surname>Sutter</surname><given-names>Esther</given-names></name></contrib><contrib contrib-type="author"><name><surname>Widmer</surname><given-names>Roland</given-names></name></contrib><contrib contrib-type="author"><name><surname>Buxton</surname><given-names>Frank</given-names></name></contrib><aff><institution>Pharma Research Department, Novartis Pharma Inc.</institution>, <addr-line>CH-4002 Basel, Switzerland</addr-line></aff></contrib-group><author-notes><corresp id="cor1"><label>*</label>To whom correspondence should be addressed at: Novartis Pharma Inc., Room K681.4.42, CH-4002 Basel, Switzerland. Tel: <phone>+41 61 69 66973</phone>; Fax: <fax>+41 61 69 66323</fax>; Email: <email>fred_am.asselbergs@pharma.novartis.com</email></corresp></author-notes><pub-date pub-type="ppub"><month>04</month><year>1998</year></pub-date><volume>26</volume><issue>7</issue><fpage>1826</fpage><lpage>1833</lpage><history><date date-type="received"><day>24</day><month>10</month><year>1997</year></date><date date-type="accepted"><day>6</day><month>2</month><year>1998</year></date></history><permissions><copyright-statement>© 1998 Oxford University Press</copyright-statement><copyright-year>1998</copyright-year></permissions><abstract><p>Two yeast artificial chromosomes containing the entire human nerve growth factor gene were isolated and mapped. By homologous recombination a luciferase gene was precisely engineered into the coding portion of the NGF gene and a neomycin selection marker was placed adjacent to one of the YAC telomeres. Expression of the YAC-based NGF reporter gene and a plasmid-based NGF reporter gene were compared with the regulation of endogenous mouse NGF protein in mouse L929 fibroblasts. In contrast to the plasmid-based reporter gene, expression and regulation of the YAC-based reporter gene was independent of the site of integration of the transgene. Basic fibroblast growth factor and okadaic acid stimulated expression of the YAC transgene, whereas transforming growth factor-β and dexamethasone inhibited it. Although cyclic AMP strongly stimulated production of the endogenous mouse NGF, no effect was seen on the human NGF reporter genes. Downregulation of the secretion of endogenous mouse NGF already occurred at an EC50 of 1–2 nM dexamethasone, but downregulation of the expression of NGF reporter genes occurred only at EC50 of 10 nM. This higher concentration was also required for upregulation of luciferase genes driven by the dexamethasone-inducible promoter of the mouse mammary tumor virus in L929 fibroblasts.</p></abstract></article-meta></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Position-independent expression of a human nerve growth factor-luciferase reporter gene cloned on a yeast artificial chromosome vector</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Position-independent expression of a human nerve growth factor-luciferase reporter gene cloned on a yeast artificial chromosome vector</title></titleInfo><name type="personal" displayLabel="corresp"><namePart type="given">Fred A.M.</namePart><namePart type="family">Asselbergs</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><affiliation>E-mail: fred_am.asselbergs@pharma.novartis.com</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Rita</namePart><namePart type="family">Grossenbacher</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Rainer</namePart><namePart type="family">Ortmann</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Bastian</namePart><namePart type="family">Hengerer</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Gary K.</namePart><namePart type="family">McMaster</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Esther</namePart><namePart type="family">Sutter</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Roland</namePart><namePart type="family">Widmer</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Frank</namePart><namePart type="family">Buxton</namePart><affiliation>Pharma Research Department, Novartis Pharma Inc., CH-4002 Basel, Switzerland</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article">research-article</genre><subject><genre>heading</genre><topic>Articles</topic></subject><originInfo><publisher>Oxford University Press</publisher><dateIssued encoding="w3cdtf">1998-04</dateIssued><copyrightDate encoding="w3cdtf">1998</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract>Two yeast artificial chromosomes containing the entire human nerve growth factor gene were isolated and mapped. By homologous recombination a luciferase gene was precisely engineered into the coding portion of the NGF gene and a neomycin selection marker was placed adjacent to one of the YAC telomeres. Expression of the YAC-based NGF reporter gene and a plasmid-based NGF reporter gene were compared with the regulation of endogenous mouse NGF protein in mouse L929 fibroblasts. In contrast to the plasmid-based reporter gene, expression and regulation of the YAC-based reporter gene was independent of the site of integration of the transgene. Basic fibroblast growth factor and okadaic acid stimulated expression of the YAC transgene, whereas transforming growth factor-β and dexamethasone inhibited it. Although cyclic AMP strongly stimulated production of the endogenous mouse NGF, no effect was seen on the human NGF reporter genes. Downregulation of the secretion of endogenous mouse NGF already occurred at an EC50 of 1–2 nM dexamethasone, but downregulation of the expression of NGF reporter genes occurred only at EC50 of 10 nM. This higher concentration was also required for upregulation of luciferase genes driven by the dexamethasone-inducible promoter of the mouse mammary tumor virus in L929 fibroblasts.</abstract><relatedItem type="host"><titleInfo><title>Nucleic Acids Research</title></titleInfo><titleInfo type="abbreviated"><title>Nucleic Acids Research</title></titleInfo><identifier type="ISSN">0305-1048</identifier><identifier type="eISSN">1362-4962</identifier><identifier type="JournalID">nar</identifier><identifier type="JournalID-hwp">nar</identifier><part><date>1998</date><detail type="volume"><caption>vol.</caption><number>26</number></detail><detail type="issue"><caption>no.</caption><number>7</number></detail><extent unit="pages"><start>1826</start><end>1833</end></extent></part></relatedItem><identifier type="istex">454B71EBFD140159884DA71C097860998154F95D</identifier><identifier type="DOI">10.1093/nar/26.7.1826</identifier><accessCondition type="use and reproduction" contentType="Copyright">© 1998 Oxford University Press</accessCondition><recordInfo><recordContentSource>OUP</recordContentSource></recordInfo></mods></metadata><enrichments><istex:catWosTEI uri="https://api.istex.fr/document/454B71EBFD140159884DA71C097860998154F95D/enrichments/catWos"><teiHeader><profileDesc><textClass><classCode scheme="WOS">BIOCHEMISTRY & MOLECULAR BIOLOGY</classCode></textClass></profileDesc></teiHeader></istex:catWosTEI></enrichments><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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