Transcriptional analysis of the isopenicillin N synthase-encoding gene of Streptomyces clavuligerus
The gene ( pcbC) encoding isopenicillin N synthase of Streptomyces clavuligerus is separated from an upstream open reading frame (ORF) by a 31-bp intergenic region. Inspection of the sequence of this intergenic region did not identify a promoter sequence. The promoter probe plasmid, pIJ4083, which c...
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| Veröffentlicht in: | Gene 1992-02, Vol.111 (1), p.77-84 |
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| creator | Petrich, Astrid K. Wu, Xiaoning Roy, Kenneth L. Jensen, Susan E. |
| description | The gene (
pcbC) encoding isopenicillin N synthase of
Streptomyces clavuligerus is separated from an upstream open reading frame (ORF) by a 31-bp intergenic region. Inspection of the sequence of this intergenic region did not identify a promoter sequence. The promoter probe plasmid, pIJ4083, which contains the promoter-less catechol-2,3-dioxygenase (C23O)-encoding gene (
xylE) as a reporter gene, was used to analyze the sequence upstream from the
pcbC gene for promoter activity. Introduction of an
SphI site at the start codon of
pcbC by site-directed mutagenesis allowed the cloning of a 335-bp fragment (−334 to + 1 in relation to the
pcbC start codon) immediately upstream from
xylE in pIJ4083. C230 activity was detected in both
Streptomyces lividans and
S. clavuligerus cultures that contained the upstream fragment, suggesting the presence of a promoter sequence. Northern analysis of total RNA extracted from
S. clavuligerus identified a monocistronic 1.2-kb transcript hybridizing to a
pcbC-specific probe. When RNA was isolated at various times during growth in liquid culture, the presence of a transcript was first detected during stationary phase. Analysis of the
pcbC transcript by primer extension located the transcription start point to a C residue within the upstream ORF, 91 bp upstream from the
pcbC start codon. |
| doi_str_mv | 10.1016/0378-1119(92)90605-O |
| format | Article |
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pcbC) encoding isopenicillin N synthase of
Streptomyces clavuligerus is separated from an upstream open reading frame (ORF) by a 31-bp intergenic region. Inspection of the sequence of this intergenic region did not identify a promoter sequence. The promoter probe plasmid, pIJ4083, which contains the promoter-less catechol-2,3-dioxygenase (C23O)-encoding gene (
xylE) as a reporter gene, was used to analyze the sequence upstream from the
pcbC gene for promoter activity. Introduction of an
SphI site at the start codon of
pcbC by site-directed mutagenesis allowed the cloning of a 335-bp fragment (−334 to + 1 in relation to the
pcbC start codon) immediately upstream from
xylE in pIJ4083. C230 activity was detected in both
Streptomyces lividans and
S. clavuligerus cultures that contained the upstream fragment, suggesting the presence of a promoter sequence. Northern analysis of total RNA extracted from
S. clavuligerus identified a monocistronic 1.2-kb transcript hybridizing to a
pcbC-specific probe. When RNA was isolated at various times during growth in liquid culture, the presence of a transcript was first detected during stationary phase. Analysis of the
pcbC transcript by primer extension located the transcription start point to a C residue within the upstream ORF, 91 bp upstream from the
pcbC start codon.</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/0378-1119(92)90605-O</identifier><identifier>PMID: 1547956</identifier><identifier>CODEN: GENED6</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>5′-primer extension ; Biological and medical sciences ; Blotting, Northern ; Catechol 1,2-Dioxygenase ; Cloning, Molecular ; Consensus Sequence ; Dioxygenases ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genes, Bacterial ; Molecular and cellular biology ; Molecular genetics ; mRNA analysis ; Oxidoreductases - genetics ; Oxidoreductases - metabolism ; Oxygenases - chemistry ; Oxygenases - metabolism ; Plasmids ; promoter probe plasmid ; Promoter Regions, Genetic ; Recombinant DNA ; Streptomyces - enzymology ; Streptomyces - genetics ; Streptomyces clavuligerus ; Transcription, Genetic ; β-lactam antibiotic biosynthesis</subject><ispartof>Gene, 1992-02, Vol.111 (1), p.77-84</ispartof><rights>1992</rights><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-7b4b9f1e8309e3a88cdb97b62a56c500792a0d8b8ffbdb52b6763b15b4b9fecc3</citedby><cites>FETCH-LOGICAL-c417t-7b4b9f1e8309e3a88cdb97b62a56c500792a0d8b8ffbdb52b6763b15b4b9fecc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0378-1119(92)90605-O$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5117388$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1547956$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Petrich, Astrid K.</creatorcontrib><creatorcontrib>Wu, Xiaoning</creatorcontrib><creatorcontrib>Roy, Kenneth L.</creatorcontrib><creatorcontrib>Jensen, Susan E.</creatorcontrib><title>Transcriptional analysis of the isopenicillin N synthase-encoding gene of Streptomyces clavuligerus</title><title>Gene</title><addtitle>Gene</addtitle><description>The gene (
pcbC) encoding isopenicillin N synthase of
Streptomyces clavuligerus is separated from an upstream open reading frame (ORF) by a 31-bp intergenic region. Inspection of the sequence of this intergenic region did not identify a promoter sequence. The promoter probe plasmid, pIJ4083, which contains the promoter-less catechol-2,3-dioxygenase (C23O)-encoding gene (
xylE) as a reporter gene, was used to analyze the sequence upstream from the
pcbC gene for promoter activity. Introduction of an
SphI site at the start codon of
pcbC by site-directed mutagenesis allowed the cloning of a 335-bp fragment (−334 to + 1 in relation to the
pcbC start codon) immediately upstream from
xylE in pIJ4083. C230 activity was detected in both
Streptomyces lividans and
S. clavuligerus cultures that contained the upstream fragment, suggesting the presence of a promoter sequence. Northern analysis of total RNA extracted from
S. clavuligerus identified a monocistronic 1.2-kb transcript hybridizing to a
pcbC-specific probe. When RNA was isolated at various times during growth in liquid culture, the presence of a transcript was first detected during stationary phase. Analysis of the
pcbC transcript by primer extension located the transcription start point to a C residue within the upstream ORF, 91 bp upstream from the
pcbC start codon.</description><subject>5′-primer extension</subject><subject>Biological and medical sciences</subject><subject>Blotting, Northern</subject><subject>Catechol 1,2-Dioxygenase</subject><subject>Cloning, Molecular</subject><subject>Consensus Sequence</subject><subject>Dioxygenases</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genes, Bacterial</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>mRNA analysis</subject><subject>Oxidoreductases - genetics</subject><subject>Oxidoreductases - metabolism</subject><subject>Oxygenases - chemistry</subject><subject>Oxygenases - metabolism</subject><subject>Plasmids</subject><subject>promoter probe plasmid</subject><subject>Promoter Regions, Genetic</subject><subject>Recombinant DNA</subject><subject>Streptomyces - enzymology</subject><subject>Streptomyces - genetics</subject><subject>Streptomyces clavuligerus</subject><subject>Transcription, Genetic</subject><subject>β-lactam antibiotic biosynthesis</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMFq3DAQhkVpSLdp36AFH0ppDk4ke2VJl0AIbRII2UPSs5Dk8UZFK7kaO7BvX212SW-Zw8xhvn8YPkK-MHrGKOvOaStkzRhTP1RzqmhHeb16RxZMClVT2sr3ZPGKfCAfEf_QUpw3x-SY8aVQvFsQ95hNRJf9OPkUTahMaVv0WKWhmp6g8phGiN75EHys7ivcxunJINQQXep9XFdriLCjH6YM45Q2WwdYuWCe5-DXkGf8RI4GExA-H-YJ-f3r5-PVTX23ur69uryr3ZKJqRZ2adXAQLZUQWukdL1VwnaN4Z3jlArVGNpLK4fB9pY3thNdaxl_iYFz7Qn5vr875vR3Bpz0xqODEEyENKNmXdNQ2bICLvegywkxw6DH7DcmbzWjeudW78TpnTitGv3iVq9K7Ovh_mw30P8P7WWW_bfD3qAzYShmncdXjDMmWikLdrHHoLh49pA1Ol9sQu8zuEn3yb_9xz_IkZf9</recordid><startdate>19920201</startdate><enddate>19920201</enddate><creator>Petrich, Astrid K.</creator><creator>Wu, Xiaoning</creator><creator>Roy, Kenneth L.</creator><creator>Jensen, Susan E.</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>7QL</scope><scope>7TM</scope><scope>C1K</scope></search><sort><creationdate>19920201</creationdate><title>Transcriptional analysis of the isopenicillin N synthase-encoding gene of Streptomyces clavuligerus</title><author>Petrich, Astrid K. ; Wu, Xiaoning ; Roy, Kenneth L. ; Jensen, Susan E.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-7b4b9f1e8309e3a88cdb97b62a56c500792a0d8b8ffbdb52b6763b15b4b9fecc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>5′-primer extension</topic><topic>Biological and medical sciences</topic><topic>Blotting, Northern</topic><topic>Catechol 1,2-Dioxygenase</topic><topic>Cloning, Molecular</topic><topic>Consensus Sequence</topic><topic>Dioxygenases</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genes, Bacterial</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>mRNA analysis</topic><topic>Oxidoreductases - genetics</topic><topic>Oxidoreductases - metabolism</topic><topic>Oxygenases - chemistry</topic><topic>Oxygenases - metabolism</topic><topic>Plasmids</topic><topic>promoter probe plasmid</topic><topic>Promoter Regions, Genetic</topic><topic>Recombinant DNA</topic><topic>Streptomyces - enzymology</topic><topic>Streptomyces - genetics</topic><topic>Streptomyces clavuligerus</topic><topic>Transcription, Genetic</topic><topic>β-lactam antibiotic biosynthesis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Petrich, Astrid K.</creatorcontrib><creatorcontrib>Wu, Xiaoning</creatorcontrib><creatorcontrib>Roy, Kenneth L.</creatorcontrib><creatorcontrib>Jensen, Susan E.</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Petrich, Astrid K.</au><au>Wu, Xiaoning</au><au>Roy, Kenneth L.</au><au>Jensen, Susan E.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transcriptional analysis of the isopenicillin N synthase-encoding gene of Streptomyces clavuligerus</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1992-02-01</date><risdate>1992</risdate><volume>111</volume><issue>1</issue><spage>77</spage><epage>84</epage><pages>77-84</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>The gene (
pcbC) encoding isopenicillin N synthase of
Streptomyces clavuligerus is separated from an upstream open reading frame (ORF) by a 31-bp intergenic region. Inspection of the sequence of this intergenic region did not identify a promoter sequence. The promoter probe plasmid, pIJ4083, which contains the promoter-less catechol-2,3-dioxygenase (C23O)-encoding gene (
xylE) as a reporter gene, was used to analyze the sequence upstream from the
pcbC gene for promoter activity. Introduction of an
SphI site at the start codon of
pcbC by site-directed mutagenesis allowed the cloning of a 335-bp fragment (−334 to + 1 in relation to the
pcbC start codon) immediately upstream from
xylE in pIJ4083. C230 activity was detected in both
Streptomyces lividans and
S. clavuligerus cultures that contained the upstream fragment, suggesting the presence of a promoter sequence. Northern analysis of total RNA extracted from
S. clavuligerus identified a monocistronic 1.2-kb transcript hybridizing to a
pcbC-specific probe. When RNA was isolated at various times during growth in liquid culture, the presence of a transcript was first detected during stationary phase. Analysis of the
pcbC transcript by primer extension located the transcription start point to a C residue within the upstream ORF, 91 bp upstream from the
pcbC start codon.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>1547956</pmid><doi>10.1016/0378-1119(92)90605-O</doi><tpages>8</tpages></addata></record> |
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| ispartof | Gene, 1992-02, Vol.111 (1), p.77-84 |
| issn | 0378-1119 1879-0038 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | 5′-primer extension Biological and medical sciences Blotting, Northern Catechol 1,2-Dioxygenase Cloning, Molecular Consensus Sequence Dioxygenases Fundamental and applied biological sciences. Psychology Gene Expression Genes, Bacterial Molecular and cellular biology Molecular genetics mRNA analysis Oxidoreductases - genetics Oxidoreductases - metabolism Oxygenases - chemistry Oxygenases - metabolism Plasmids promoter probe plasmid Promoter Regions, Genetic Recombinant DNA Streptomyces - enzymology Streptomyces - genetics Streptomyces clavuligerus Transcription, Genetic β-lactam antibiotic biosynthesis |
| title | Transcriptional analysis of the isopenicillin N synthase-encoding gene of Streptomyces clavuligerus |
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