Controlled expression of heterologous cytochrome P450e cDNA in Saccharomyces cerevisiae. I. Construction and expression of a complete rat cytochrome P450e cDNA
A complete rat cytochrome P450e cDNA was constructed and placed between the yeast-inducible acid phosphatase (PHO5) promoter and terminator. This expression cassette was inserted into the yeast-replicating vector pDP34 which contains the entire yeast 2 μ plasmid element, the dLEU2 and the URA3 marke...
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| Veröffentlicht in: | Journal of biotechnology 1989, Vol.9 (4), p.255-272 |
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| container_issue | 4 |
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| container_title | Journal of biotechnology |
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| creator | Zurbriggen, Beat Böhlen, Elizabeth Sanglard, Dominique Käppeli, Othmar Fiechter, Armin |
| description | A complete rat cytochrome P450e cDNA was constructed and placed between the yeast-inducible acid phosphatase (PHO5) promoter and terminator. This expression cassette was inserted into the yeast-replicating vector pDP34 which contains the entire yeast 2 μ plasmid element, the dLEU2 and the URA3 marker genes.
Saccharomyces cerevisiae was used as a recipient for this construction and was grown under phosphate limitation (induced conditions) in shake flask cultures. Cytochrome P450e production was measured by CO-difference spectra. Cytochrome P450e-specific transcripts were detected by Northern blot analysis and cytochrome P450e proteins by immunoblotting with monoclonal cytochrome P450b antibodies. The intact cytochrome P450e proteins were localized in microsomes. In
S. cerevisiae, the cytochrome P450e proteins have been shown to be functional and coupled with yeast NADPH-P450 reductase to form a monooxygenase system. |
| doi_str_mv | 10.1016/0168-1656(89)90002-3 |
| format | Article |
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Saccharomyces cerevisiae was used as a recipient for this construction and was grown under phosphate limitation (induced conditions) in shake flask cultures. Cytochrome P450e production was measured by CO-difference spectra. Cytochrome P450e-specific transcripts were detected by Northern blot analysis and cytochrome P450e proteins by immunoblotting with monoclonal cytochrome P450b antibodies. The intact cytochrome P450e proteins were localized in microsomes. In
S. cerevisiae, the cytochrome P450e proteins have been shown to be functional and coupled with yeast NADPH-P450 reductase to form a monooxygenase system.</description><identifier>ISSN: 0168-1656</identifier><identifier>EISSN: 1873-4863</identifier><identifier>DOI: 10.1016/0168-1656(89)90002-3</identifier><language>eng</language><publisher>Elsevier B.V</publisher><subject>cDNA ; cloning ; cloning vectors ; Complete cDNA ; Cytochrome P450e ; Deleted cDNA ; PHO5 expression system ; plasmids ; Saccharomyces cerevisiae</subject><ispartof>Journal of biotechnology, 1989, Vol.9 (4), p.255-272</ispartof><rights>1989</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c286t-eb779f0482afaee9c69504231fbe4cc39db50ece50cdb5b490bc99ffe4f3bf993</citedby><cites>FETCH-LOGICAL-c286t-eb779f0482afaee9c69504231fbe4cc39db50ece50cdb5b490bc99ffe4f3bf993</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0168-1656(89)90002-3$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,4024,27923,27924,27925,45995</link.rule.ids></links><search><creatorcontrib>Zurbriggen, Beat</creatorcontrib><creatorcontrib>Böhlen, Elizabeth</creatorcontrib><creatorcontrib>Sanglard, Dominique</creatorcontrib><creatorcontrib>Käppeli, Othmar</creatorcontrib><creatorcontrib>Fiechter, Armin</creatorcontrib><title>Controlled expression of heterologous cytochrome P450e cDNA in Saccharomyces cerevisiae. I. Construction and expression of a complete rat cytochrome P450e cDNA</title><title>Journal of biotechnology</title><description>A complete rat cytochrome P450e cDNA was constructed and placed between the yeast-inducible acid phosphatase (PHO5) promoter and terminator. This expression cassette was inserted into the yeast-replicating vector pDP34 which contains the entire yeast 2 μ plasmid element, the dLEU2 and the URA3 marker genes.
Saccharomyces cerevisiae was used as a recipient for this construction and was grown under phosphate limitation (induced conditions) in shake flask cultures. Cytochrome P450e production was measured by CO-difference spectra. Cytochrome P450e-specific transcripts were detected by Northern blot analysis and cytochrome P450e proteins by immunoblotting with monoclonal cytochrome P450b antibodies. The intact cytochrome P450e proteins were localized in microsomes. In
S. cerevisiae, the cytochrome P450e proteins have been shown to be functional and coupled with yeast NADPH-P450 reductase to form a monooxygenase system.</description><subject>cDNA</subject><subject>cloning</subject><subject>cloning vectors</subject><subject>Complete cDNA</subject><subject>Cytochrome P450e</subject><subject>Deleted cDNA</subject><subject>PHO5 expression system</subject><subject>plasmids</subject><subject>Saccharomyces cerevisiae</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><recordid>eNp9kc9KAzEQxoMoWKtv4CEn0cPWZDe73VwEqf8KooJ6DtnpxEa2m5qkxT6Nr2pqxYOIhyED883vI98QcsjZgDNenaaqM16V1XEtTyRjLM-KLdLj9bDIRF0V26T3I9kleyG8Jo2QJe-Rj5HrondtixOK73OPIVjXUWfoFCOmgXtxi0BhFR1MvZshfRAlQwoXd-fUdvRRA0x1GqwAkww9Lm2wGgd0PKCJHaJfQFwjdffbQVNws3mbfKjX8W-PfbJjdBvw4Pvtk-ery6fRTXZ7fz0end9mkNdVzLAZDqVhos610YgSKlkykRfcNCgACjlpEhGwZJC6RkjWgJTGoDBFY6Qs-uRow51797bAENXMBsC21R2mABQvc554VRKKjRC8C8GjUXNvZ9qvFGdqfQ21jlqto1a1VF_XUEVaO9usYfrE0qJXASx2gBPrEaKaOPs_4BO7sZRR</recordid><startdate>1989</startdate><enddate>1989</enddate><creator>Zurbriggen, Beat</creator><creator>Böhlen, Elizabeth</creator><creator>Sanglard, Dominique</creator><creator>Käppeli, Othmar</creator><creator>Fiechter, Armin</creator><general>Elsevier B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>1989</creationdate><title>Controlled expression of heterologous cytochrome P450e cDNA in Saccharomyces cerevisiae. I. Construction and expression of a complete rat cytochrome P450e cDNA</title><author>Zurbriggen, Beat ; Böhlen, Elizabeth ; Sanglard, Dominique ; Käppeli, Othmar ; Fiechter, Armin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c286t-eb779f0482afaee9c69504231fbe4cc39db50ece50cdb5b490bc99ffe4f3bf993</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>cDNA</topic><topic>cloning</topic><topic>cloning vectors</topic><topic>Complete cDNA</topic><topic>Cytochrome P450e</topic><topic>Deleted cDNA</topic><topic>PHO5 expression system</topic><topic>plasmids</topic><topic>Saccharomyces cerevisiae</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zurbriggen, Beat</creatorcontrib><creatorcontrib>Böhlen, Elizabeth</creatorcontrib><creatorcontrib>Sanglard, Dominique</creatorcontrib><creatorcontrib>Käppeli, Othmar</creatorcontrib><creatorcontrib>Fiechter, Armin</creatorcontrib><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zurbriggen, Beat</au><au>Böhlen, Elizabeth</au><au>Sanglard, Dominique</au><au>Käppeli, Othmar</au><au>Fiechter, Armin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Controlled expression of heterologous cytochrome P450e cDNA in Saccharomyces cerevisiae. I. Construction and expression of a complete rat cytochrome P450e cDNA</atitle><jtitle>Journal of biotechnology</jtitle><date>1989</date><risdate>1989</risdate><volume>9</volume><issue>4</issue><spage>255</spage><epage>272</epage><pages>255-272</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><abstract>A complete rat cytochrome P450e cDNA was constructed and placed between the yeast-inducible acid phosphatase (PHO5) promoter and terminator. This expression cassette was inserted into the yeast-replicating vector pDP34 which contains the entire yeast 2 μ plasmid element, the dLEU2 and the URA3 marker genes.
Saccharomyces cerevisiae was used as a recipient for this construction and was grown under phosphate limitation (induced conditions) in shake flask cultures. Cytochrome P450e production was measured by CO-difference spectra. Cytochrome P450e-specific transcripts were detected by Northern blot analysis and cytochrome P450e proteins by immunoblotting with monoclonal cytochrome P450b antibodies. The intact cytochrome P450e proteins were localized in microsomes. In
S. cerevisiae, the cytochrome P450e proteins have been shown to be functional and coupled with yeast NADPH-P450 reductase to form a monooxygenase system.</abstract><pub>Elsevier B.V</pub><doi>10.1016/0168-1656(89)90002-3</doi><tpages>18</tpages></addata></record> |
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| ispartof | Journal of biotechnology, 1989, Vol.9 (4), p.255-272 |
| issn | 0168-1656 1873-4863 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_15214236 |
| source | Access via ScienceDirect (Elsevier) |
| subjects | cDNA cloning cloning vectors Complete cDNA Cytochrome P450e Deleted cDNA PHO5 expression system plasmids Saccharomyces cerevisiae |
| title | Controlled expression of heterologous cytochrome P450e cDNA in Saccharomyces cerevisiae. I. Construction and expression of a complete rat cytochrome P450e cDNA |
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