Tightly regulated vectors for the cloning and expression of toxic genes
A series of low-copy expression vectors that permits the stable maintenance and regulated expression of highly toxic gene products has been developed. These vectors utilize the lactose promoter/operator system, and protect against read-through transcription from other promoters on the plasmid by pla...
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| Veröffentlicht in: | Journal of microbiological methods 2004-08, Vol.58 (2), p.243-250 |
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| container_title | Journal of microbiological methods |
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| creator | Anthony, Larry C Suzuki, Hideki Filutowicz, Marcin |
| description | A series of low-copy expression vectors that permits the stable maintenance and regulated expression of highly toxic gene products has been developed. These vectors utilize the lactose promoter/operator system, and protect against read-through transcription from other promoters on the plasmid by placement of the
rrnB T1T2 terminators upstream of the lactose promoter. For additional regulatory control, the vectors utilize low-copy origins of replication. Either the pMPP6 origin (pSC101-derived) is used for cloning into
Escherichia coli or related species, or the broad-host-range RK2 origin of replication is utilized for cloning into the majority of Gram-negative bacteria. The resulting plasmids have no detectable leaky expression. To test these vectors, the genes for the bacteriocidal colicins D, E3, and E7 were cloned and stably maintained in the absence of their immunity genes. Upon induction with isopropyl-β-
d-thiogalactopyranoside (IPTG), cell death was observed, indicating expression of each colicin. These low-copy expression vectors will be useful for the cloning and expression of toxic genes in bacterial systems. |
| doi_str_mv | 10.1016/j.mimet.2004.04.003 |
| format | Article |
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rrnB T1T2 terminators upstream of the lactose promoter. For additional regulatory control, the vectors utilize low-copy origins of replication. Either the pMPP6 origin (pSC101-derived) is used for cloning into
Escherichia coli or related species, or the broad-host-range RK2 origin of replication is utilized for cloning into the majority of Gram-negative bacteria. The resulting plasmids have no detectable leaky expression. To test these vectors, the genes for the bacteriocidal colicins D, E3, and E7 were cloned and stably maintained in the absence of their immunity genes. Upon induction with isopropyl-β-
d-thiogalactopyranoside (IPTG), cell death was observed, indicating expression of each colicin. These low-copy expression vectors will be useful for the cloning and expression of toxic genes in bacterial systems.</description><subject>Bacteriological Techniques</subject><subject>Biological and medical sciences</subject><subject>Cloning, Molecular</subject><subject>Colicins - genetics</subject><subject>Colicins - toxicity</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genes, Bacterial</subject><subject>Genetic Techniques</subject><subject>Genetic Vectors</subject><subject>Lac Operon</subject><subject>Microbiology</subject><subject>Plasmids - genetics</subject><subject>Promoter Regions, Genetic</subject><issn>0167-7012</issn><issn>1872-8359</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1LAzEQhoMotlZ_gSC56G1rPja7m4MHKVqFgpd6Dml20qbsbmqyLe2_d9cW9CQMzOV5X2YehG4pGVNCs8f1uHY1tGNGSDruh_AzNKRFzpKCC3mOhh2VJzmhbICuYlwTQgVPi0s0oILxVDA2RNO5W67a6oADLLeVbqHEOzCtDxFbH3C7Amwq37hmiXVTYthvAsTofIO9xa3fO4OX0EC8RhdWVxFuTnuEPl9f5pO3ZPYxfZ88zxKTFqxNeCYMkxnN86LMoISFNMIQXpRUUrBSWplxIGmhBRCQgqaZNgvgxlhhmQXGR-jh2LsJ_msLsVW1iwaqSjfgt1HRons6Y7wD-RE0wccYwKpNcLUOB0WJ6v2ptfrxp3p_qh_Sp-5O9dtFDeVv5iSsA-5PgI5GVzboxrj4h5Mpy2necU9HDjoZOwdBReOgMVC60PlVpXf_HvINHpaPlw</recordid><startdate>20040801</startdate><enddate>20040801</enddate><creator>Anthony, Larry C</creator><creator>Suzuki, Hideki</creator><creator>Filutowicz, Marcin</creator><general>Elsevier B.V</general><general>Elsevier Science</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>7QO</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20040801</creationdate><title>Tightly regulated vectors for the cloning and expression of toxic genes</title><author>Anthony, Larry C ; Suzuki, Hideki ; Filutowicz, Marcin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c482t-365c2961778d6edeb9c5c038d191ef99f963e048a5e0e95146acbe3ccf5f2fe23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Bacteriological Techniques</topic><topic>Biological and medical sciences</topic><topic>Cloning, Molecular</topic><topic>Colicins - genetics</topic><topic>Colicins - toxicity</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genes, Bacterial</topic><topic>Genetic Techniques</topic><topic>Genetic Vectors</topic><topic>Lac Operon</topic><topic>Microbiology</topic><topic>Plasmids - genetics</topic><topic>Promoter Regions, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Anthony, Larry C</creatorcontrib><creatorcontrib>Suzuki, Hideki</creatorcontrib><creatorcontrib>Filutowicz, Marcin</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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of microbiological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Anthony, Larry C</au><au>Suzuki, Hideki</au><au>Filutowicz, Marcin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Tightly regulated vectors for the cloning and expression of toxic genes</atitle><jtitle>Journal of microbiological methods</jtitle><addtitle>J Microbiol Methods</addtitle><date>2004-08-01</date><risdate>2004</risdate><volume>58</volume><issue>2</issue><spage>243</spage><epage>250</epage><pages>243-250</pages><issn>0167-7012</issn><eissn>1872-8359</eissn><coden>JMIMDQ</coden><abstract>A series of low-copy expression vectors that permits the stable maintenance and regulated expression of highly toxic gene products has been developed. These vectors utilize the lactose promoter/operator system, and protect against read-through transcription from other promoters on the plasmid by placement of the
rrnB T1T2 terminators upstream of the lactose promoter. For additional regulatory control, the vectors utilize low-copy origins of replication. Either the pMPP6 origin (pSC101-derived) is used for cloning into
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| ispartof | Journal of microbiological methods, 2004-08, Vol.58 (2), p.243-250 |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Bacteriological Techniques Biological and medical sciences Cloning, Molecular Colicins - genetics Colicins - toxicity Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Expression Genes, Bacterial Genetic Techniques Genetic Vectors Lac Operon Microbiology Plasmids - genetics Promoter Regions, Genetic |
| title | Tightly regulated vectors for the cloning and expression of toxic genes |
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