Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter
Identifieur interne : 001D17 ( Istex/Corpus ); précédent : 001D16; suivant : 001D18Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter
Auteurs : Takekoshi Masataka ; Maeda-Takekoshi Fumiko ; Ihara Seiji ; Sakuma Sadatoshi ; Watanabe YasushiSource :
- Gene [ 0378-1119 ] ; 1991.
English descriptors
- Teeft :
- Bgal, Bgal activity, Blue plaques, Chimeric, Chimeric gene, Chimeric iucz gene, Cytomegalovirus, Early stage, Foreign gene, Foreign genes, Fragment, Gene, Genome, Genome structures, Hcmv, Hcmv genome, Hcmvlacr, Iacz gene, Insertion, Iucz, Iucz gene, Late glycoprotein, Late stages, Marker genes, Mocarski, Negative regulation, Open circles, Opposite directions, Plasmid, Progeny genome, Promoter, Recombinant, Recombinant genomes, Recombinant virus, Recombinant viruses, Reversion rate, Simian virus, Viral, Viral proteins, Virion dnas, Virus replication, Virus vector.
Abstract
Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.
Url:
DOI: 10.1016/0378-1119(91)90413-6
Links to Exploration step
ISTEX:921795161D969F4C358BC51E4BF2CCDB6D35DFEALe document en format XML
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(0465)37-3661</mods:affiliation></affiliation></author><author><name sortKey="Yasushi, Watanabe" sort="Yasushi, Watanabe" uniqKey="Yasushi W" first="Watanabe" last="Yasushi">Watanabe Yasushi</name><affiliation><mods:affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Gene</title><title level="j" type="abbrev">GENE</title><idno type="ISSN">0378-1119</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1991">1991</date><biblScope unit="volume">101</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="209">209</biblScope><biblScope unit="page" to="213">213</biblScope></imprint><idno type="ISSN">0378-1119</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0378-1119</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Bgal</term><term>Bgal activity</term><term>Blue plaques</term><term>Chimeric</term><term>Chimeric gene</term><term>Chimeric iucz gene</term><term>Cytomegalovirus</term><term>Early stage</term><term>Foreign gene</term><term>Foreign genes</term><term>Fragment</term><term>Gene</term><term>Genome</term><term>Genome structures</term><term>Hcmv</term><term>Hcmv genome</term><term>Hcmvlacr</term><term>Iacz gene</term><term>Insertion</term><term>Iucz</term><term>Iucz gene</term><term>Late glycoprotein</term><term>Late stages</term><term>Marker genes</term><term>Mocarski</term><term>Negative regulation</term><term>Open circles</term><term>Opposite directions</term><term>Plasmid</term><term>Progeny genome</term><term>Promoter</term><term>Recombinant</term><term>Recombinant genomes</term><term>Recombinant virus</term><term>Recombinant viruses</term><term>Reversion rate</term><term>Simian virus</term><term>Viral</term><term>Viral proteins</term><term>Virion dnas</term><term>Virus replication</term><term>Virus vector</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>genome</json:string><json:string>recombinant</json:string><json:string>hcmv</json:string><json:string>viral</json:string><json:string>chimeric</json:string><json:string>iucz</json:string><json:string>cytomegalovirus</json:string><json:string>plasmid</json:string><json:string>bgal</json:string><json:string>hcmvlacr</json:string><json:string>mocarski</json:string><json:string>recombinant viruses</json:string><json:string>hcmv genome</json:string><json:string>insertion</json:string><json:string>iucz gene</json:string><json:string>promoter</json:string><json:string>fragment</json:string><json:string>late glycoprotein</json:string><json:string>late stages</json:string><json:string>bgal activity</json:string><json:string>recombinant genomes</json:string><json:string>foreign gene</json:string><json:string>open circles</json:string><json:string>iacz gene</json:string><json:string>chimeric gene</json:string><json:string>reversion rate</json:string><json:string>recombinant virus</json:string><json:string>blue plaques</json:string><json:string>virus replication</json:string><json:string>marker genes</json:string><json:string>viral proteins</json:string><json:string>opposite directions</json:string><json:string>virion dnas</json:string><json:string>chimeric iucz gene</json:string><json:string>genome structures</json:string><json:string>progeny genome</json:string><json:string>early stage</json:string><json:string>simian virus</json:string><json:string>foreign genes</json:string><json:string>virus vector</json:string><json:string>negative regulation</json:string><json:string>gene</json:string></teeft></keywords><author><json:item><name>Takekoshi Masataka</name><affiliations><json:string>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</json:string></affiliations></json:item><json:item><name>Maeda-Takekoshi Fumiko</name><affiliations><json:string>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</json:string></affiliations></json:item><json:item><name>Ihara Seiji</name><affiliations><json:string>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</json:string></affiliations></json:item><json:item><name>Sakuma Sadatoshi</name><affiliations><json:string>Molecular Oncology Division, Meiji Institute of Health Science, 540 Naruda, Odawara 250 Japan Tel. (0465)37-3661</json:string></affiliations></json:item><json:item><name>Watanabe Yasushi</name><affiliations><json:string>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Recombinant DNA</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>animal virus vector</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>live vaccine</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>replacement insertion</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>homologous recombination</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>lacZ</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>AraC : cytosine arabinoside</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>βGal : β-galactosidase</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>bp : base pair(s)</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>HCMV : human cytomegalovirus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>HEL : human embryonic lung</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>kb : kilobase(s) or 1000 bp</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>mAb : monoclonal antibody</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>MCS : multiple cloning site</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>MEM : Eagle's minimum essential medium</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>moi : multiplicity of infection</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>nt : nucleotide(s)</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>pfu : plaque-forming unit(s)</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Pollk : Klenow (large) fragment of E. coll DNA polymerase I</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>SDS : sodium dodecyl sulfate</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>SV40 : simian virus 40</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>XGal : 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside</value></json:item></subject><arkIstex>ark:/67375/6H6-28681FFT-1</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.</abstract><qualityIndicators><score>6.353</score><pdfWordCount>2493</pdfWordCount><pdfCharCount>17841</pdfCharCount><pdfVersion>1.4</pdfVersion><pdfPageCount>5</pdfPageCount><pdfPageSize>612 x 792 pts (letter)</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>155</abstractWordCount><abstractCharCount>1013</abstractCharCount><keywordCount>22</keywordCount></qualityIndicators><title>Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter</title><pmid><json:string>1647360</json:string></pmid><pii><json:string>0378-1119(91)90413-6</json:string></pii><genre><json:string>research-article</json:string></genre><serie><title>Proc. Natl. Acad. Sci. USA</title><language><json:string>unknown</json:string></language><volume>82</volume><pages><first>4773</first><last>4777</last></pages></serie><host><title>Gene</title><language><json:string>unknown</json:string></language><publicationDate>1991</publicationDate><issn><json:string>0378-1119</json:string></issn><pii><json:string>S0378-1119(00)X0407-6</json:string></pii><volume>101</volume><issue>2</issue><pages><first>209</first><last>213</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1991</json:string></date><geogName></geogName><orgName><json:string>Research Aid</json:string><json:string>Department of Molecular Tokai University School of Medicine, Bohseidai, Isehara Tel</json:string><json:string>Ministry of Education, Science and Culture</json:string><json:string>Tokai University</json:string><json:string>Meiji Institute of Health Science</json:string><json:string>Chemicon International, Inc.</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Furukawa</json:string><json:string>K. Matsubara</json:string><json:string>J. Gen</json:string><json:string>M. Takekoshi</json:string></persName><placeName><json:string>Mannheim</json:string><json:string>HEL</json:string><json:string>Japan</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Compels and Minison (1989)</json:string><json:string>Mackett and Smith, 1986</json:string><json:string>Manning and Mocarski, 1988</json:string><json:string>Lowe et al., 1987</json:string><json:string>Roizman and Jenkins, 1985</json:string><json:string>Spaete and Mocarski (1987)</json:string><json:string>Miller, 1972</json:string><json:string>Jenkins et al., 1985</json:string><json:string>Shirakawa et al., 1984</json:string><json:string>Ihara et al., 1986</json:string><json:string>Rigby, 1983</json:string><json:string>Sauer et al., 1987</json:string><json:string>Stinski, 1978</json:string><json:string>Smith and Moss, 1983</json:string><json:string>Westrate et al., 1980</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-28681FFT-1</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - genetics & heredity</json:string></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - developmental biology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - Genetics</json:string><json:string>1 - Health Sciences</json:string><json:string>2 - Medicine</json:string><json:string>3 - General Medicine</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences biologiques fondamentales et appliquees. psychologie</json:string><json:string>4 - microbiologie</json:string></inist></categories><publicationDate>1991</publicationDate><copyrightDate>1991</copyrightDate><doi><json:string>10.1016/0378-1119(91)90413-6</json:string></doi><id>921795161D969F4C358BC51E4BF2CCDB6D35DFEA</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-28681FFT-1/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-28681FFT-1/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/6H6-28681FFT-1/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a">Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>elsevier</p></licence></availability><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M"></p><date>1991</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter</title><author xml:id="author-0000"><persName><forename type="first">Takekoshi</forename><surname>Masataka</surname></persName><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Maeda-Takekoshi</forename><surname>Fumiko</surname></persName><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Ihara</forename><surname>Seiji</surname></persName><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation></author><author xml:id="author-0003"><persName><forename type="first">Sakuma</forename><surname>Sadatoshi</surname></persName><affiliation>Molecular Oncology Division, Meiji Institute of Health Science, 540 Naruda, Odawara 250 Japan Tel. (0465)37-3661</affiliation></author><author xml:id="author-0004"><persName><forename type="first">Watanabe</forename><surname>Yasushi</surname></persName><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation></author><idno type="istex">921795161D969F4C358BC51E4BF2CCDB6D35DFEA</idno><idno type="ark">ark:/67375/6H6-28681FFT-1</idno><idno type="DOI">10.1016/0378-1119(91)90413-6</idno><idno type="PII">0378-1119(91)90413-6</idno></analytic><monogr><title level="j">Gene</title><title level="j" type="abbrev">GENE</title><idno type="pISSN">0378-1119</idno><idno type="PII">S0378-1119(00)X0407-6</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1991"></date><biblScope unit="volume">101</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="209">209</biblScope><biblScope unit="page" to="213">213</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1991</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>Recombinant DNA</term></item><item><term>animal virus vector</term></item><item><term>live vaccine</term></item><item><term>replacement insertion</term></item><item><term>homologous recombination</term></item><item><term>lacZ</term></item></list></keywords></textClass><textClass><keywords scheme="keyword"><list><head>Abbreviations</head><item><term>AraC</term><term>cytosine arabinoside</term></item><item><term>βGal</term><term>β-galactosidase</term></item><item><term>bp</term><term>base pair(s)</term></item><item><term>HCMV</term><term>human cytomegalovirus</term></item><item><term>HEL</term><term>human embryonic lung</term></item><item><term>kb</term><term>kilobase(s) or 1000 bp</term></item><item><term>mAb</term><term>monoclonal antibody</term></item><item><term>MCS</term><term>multiple cloning site</term></item><item><term>MEM</term><term>Eagle's minimum essential medium</term></item><item><term>moi</term><term>multiplicity of infection</term></item><item><term>nt</term><term>nucleotide(s)</term></item><item><term>pfu</term><term>plaque-forming unit(s)</term></item><item><term>Pollk</term><term>Klenow (large) fragment of E. coll DNA polymerase I</term></item><item><term>SDS</term><term>sodium dodecyl sulfate</term></item><item><term>SV40</term><term>simian virus 40</term></item><item><term>XGal</term><term>5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1990-12-04">Modified</change><change when="1991">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-28681FFT-1/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>GENE</jid><aid>91904136</aid><ce:pii>0378-1119(91)90413-6</ce:pii><ce:doi>10.1016/0378-1119(91)90413-6</ce:doi><ce:copyright type="unknown" year="1991"></ce:copyright></item-info><head><ce:title>Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter</ce:title><ce:author-group><ce:author><ce:surname>Masataka</ce:surname><ce:given-name>Takekoshi</ce:given-name><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:surname>Fumiko</ce:surname><ce:given-name>Maeda-Takekoshi</ce:given-name><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:surname>Seiji</ce:surname><ce:given-name>Ihara</ce:given-name><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:surname>Sadatoshi</ce:surname><ce:given-name>Sakuma</ce:given-name><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:surname>Yasushi</ce:surname><ce:given-name>Watanabe</ce:given-name><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>Molecular Oncology Division, Meiji Institute of Health Science, 540 Naruda, Odawara 250 Japan Tel. (0465)37-3661</ce:textfn></ce:affiliation></ce:author-group><ce:date-received day="18" month="9" year="1990"></ce:date-received><ce:date-revised day="4" month="12" year="1990"></ce:date-revised><ce:date-accepted day="9" month="12" year="1990"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>On the basis of a previous finding that the 7.8-kb <ce:italic>Hind</ce:italic>III-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric <ce:italic>lacZ</ce:italic> gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric <ce:italic>lacZ</ce:italic> gene introduced was stable and no <ce:italic>lacZ</ce:italic><ce:sup>−</ce:sup> revertants have been observed so far.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Recombinant DNA</ce:text></ce:keyword><ce:keyword><ce:text>animal virus vector</ce:text></ce:keyword><ce:keyword><ce:text>live vaccine</ce:text></ce:keyword><ce:keyword><ce:text>replacement insertion</ce:text></ce:keyword><ce:keyword><ce:text>homologous recombination</ce:text></ce:keyword><ce:keyword><ce:text><ce:italic>lacZ</ce:italic></ce:text></ce:keyword></ce:keywords><ce:keywords class="abr"><ce:section-title>Abbreviations</ce:section-title><ce:keyword><ce:text>AraC</ce:text><ce:keyword><ce:text>cytosine arabinoside</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>βGal</ce:text><ce:keyword><ce:text>β-galactosidase</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>bp</ce:text><ce:keyword><ce:text>base pair(s)</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>HCMV</ce:text><ce:keyword><ce:text>human cytomegalovirus</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>HEL</ce:text><ce:keyword><ce:text>human embryonic lung</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>kb</ce:text><ce:keyword><ce:text>kilobase(s) or 1000 bp</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>mAb</ce:text><ce:keyword><ce:text>monoclonal antibody</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>MCS</ce:text><ce:keyword><ce:text>multiple cloning site</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>MEM</ce:text><ce:keyword><ce:text>Eagle's minimum essential medium</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>moi</ce:text><ce:keyword><ce:text>multiplicity of infection</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>nt</ce:text><ce:keyword><ce:text>nucleotide(s)</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>pfu</ce:text><ce:keyword><ce:text>plaque-forming unit(s)</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>Pollk</ce:text><ce:keyword><ce:text>Klenow (large) fragment of <ce:italic>E. coll</ce:italic> DNA polymerase I</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>SDS</ce:text><ce:keyword><ce:text>sodium dodecyl sulfate</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>SV40</ce:text><ce:keyword><ce:text>simian virus 40</ce:text></ce:keyword></ce:keyword><ce:keyword><ce:text>XGal</ce:text><ce:keyword><ce:text>5-bromo-4-chloro-3-indolyl-β-<ce:small-caps>d</ce:small-caps>-galactopyranoside</ce:text></ce:keyword></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter</title></titleInfo><name type="personal"><namePart type="given">Takekoshi</namePart><namePart type="family">Masataka</namePart><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Maeda-Takekoshi</namePart><namePart type="family">Fumiko</namePart><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ihara</namePart><namePart type="family">Seiji</namePart><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Sakuma</namePart><namePart type="family">Sadatoshi</namePart><affiliation>Molecular Oncology Division, Meiji Institute of Health Science, 540 Naruda, Odawara 250 Japan Tel. (0465)37-3661</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Watanabe</namePart><namePart type="family">Yasushi</namePart><affiliation>Department of Molecular Biology, Tokai University School of Medicine, Bohseidai, Isehara 259-11 Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1991</dateIssued><dateModified encoding="w3cdtf">1990-12-04</dateModified><copyrightDate encoding="w3cdtf">1991</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: On the basis of a previous finding that the 7.8-kb HindIII-O fragment of the human cytomegalovirus strain Towne genome is nonessential for viral replication, we constructed a vector, pKM, that directs introduction of foreign genes by homologous recombination precisely replacing the O fragment. Using this vector, we constructed Towne-strain-derived recombinant viruses in which a chimeric lacZ gene fused to the simian virus 40 promoter and a poly(A) signal were inserted in place of the O fragment. Two types of recombinants were obtained which carried the chimeric gene in opposite directions. β-Galacto- sidase (βGal) was produced throughout the infection cycle in human embryonic lung cells infected with these recombinants, and the rate of its synthesis in the early stages of infection was comparable to that of synthesis of a 65-kDa viral glycoprotein, one of the abundantly produced viral proteins. The chimeric lacZ gene introduced was stable and no lacZ− revertants have been observed so far.</abstract><subject><genre>Keywords</genre><topic>Recombinant DNA</topic><topic>animal virus vector</topic><topic>live vaccine</topic><topic>replacement insertion</topic><topic>homologous recombination</topic><topic>lacZ</topic></subject><subject><genre>Abbreviations</genre><topic>AraC : cytosine arabinoside</topic><topic>βGal : β-galactosidase</topic><topic>bp : base pair(s)</topic><topic>HCMV : human cytomegalovirus</topic><topic>HEL : human embryonic lung</topic><topic>kb : kilobase(s) or 1000 bp</topic><topic>mAb : monoclonal antibody</topic><topic>MCS : multiple cloning site</topic><topic>MEM : Eagle's minimum essential medium</topic><topic>moi : multiplicity of infection</topic><topic>nt : nucleotide(s)</topic><topic>pfu : plaque-forming unit(s)</topic><topic>Pollk : Klenow (large) fragment of E. coll DNA polymerase I</topic><topic>SDS : sodium dodecyl sulfate</topic><topic>SV40 : simian virus 40</topic><topic>XGal : 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside</topic></subject><relatedItem type="host"><titleInfo><title>Gene</title></titleInfo><titleInfo type="abbreviated"><title>GENE</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1991</dateIssued></originInfo><identifier type="ISSN">0378-1119</identifier><identifier type="PII">S0378-1119(00)X0407-6</identifier><part><date>1991</date><detail type="volume"><number>101</number><caption>vol.</caption></detail><detail type="issue"><number>2</number><caption>no.</caption></detail><extent unit="issue-pages"><start>165</start><end>311</end></extent><extent unit="pages"><start>209</start><end>213</end></extent></part></relatedItem><identifier type="istex">921795161D969F4C358BC51E4BF2CCDB6D35DFEA</identifier><identifier type="ark">ark:/67375/6H6-28681FFT-1</identifier><identifier type="DOI">10.1016/0378-1119(91)90413-6</identifier><identifier type="PII">0378-1119(91)90413-6</identifier><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-28681FFT-1/record.json</uri></json:item></metadata></istex></record>Pour manipuler ce document sous Unix (Dilib)
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