| A series of wide-host-range low-copy-number vectors that allow direct screening for recombinants. | |
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MedLine Citation:
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PMID: 1847347 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A series of controlled expression vectors was constructed based on the wide-host-range plasmid pMMB66EH. Some of these new vectors code for the alpha-peptide of beta-galactosidase and allow the direct screening of recombinant clones by inactivation of alpha-complementation. The bla gene was replaced in some plasmids by the cat gene of Tn9 coding for chloramphenicol resistance, extending the use into beta-lactam-resistant strains. They all feature either the tac or taclac (tac-lac UV5 in tandem) promoters in front of a polylinker followed by the rrnB transcriptional stop point. These vectors were tested by subcloning the xylE gene coding for the Pseudomonas putida catechol 2,3-oxygenase and the Escherichia coli lamB gene coding for the lambda receptor. The expression of these genes in E. coli indicated that the tac promoter is five times stronger than the taclac promoter and that both were tightly regulated. The tac promoter in Pseudomonas syringae pv glycinea and Xanthomonas campestris pv vesicatoria had a strength similar to that in E. coli, while the taclac promoter was much weaker, reaching only 6.5 and 3% of the level of expression of the tac promoter, respectively. The taclac promoter, however, proved to be useful for the cloning in E. coli of DNA fragments that were unstable in vectors with stronger promoters and higher copy number. Expression of the lamB gene in Vibrio cholerae strain TRH7000 was not sufficient to permit cosmid transduction. Two subunits of the E. coli mannose permease, coded by the ptsP and ptsM genes, are also required for cosmid DNA penetration into the recipient cells. |
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Authors:
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V M Morales; A Bäckman; M Bagdasarian |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Gene Volume: 97 ISSN: 0378-1119 ISO Abbreviation: Gene Publication Date: 1991 Jan |
Date Detail:
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Created Date: 1991-03-27 Completed Date: 1991-03-27 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 7706761 Medline TA: Gene Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 39-47 Citation Subset: IM |
Affiliation:
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Unit for Applied Cell and Molecular Biology, Umeå University, Sweden. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/M37846; M37847; M37848; M37849; M37850; M37851; M37852; M63445; M74185; S75878 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Base Sequence Chloramphenicol Resistance / genetics Cloning, Molecular Cosmids DNA Transposable Elements DNA, Recombinant / genetics* Gene Expression Regulation, Bacterial Genes, Bacterial Genetic Complementation Test Genetic Vectors* Lac Operon Molecular Sequence Data Phosphoenolpyruvate Sugar Phosphotransferase System / genetics Plasmids* Promoter Regions, Genetic Restriction Mapping Transcription, Genetic Transduction, Genetic Vibrio cholerae / genetics |
| Chemical | |
Reg. No./Substance:
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0/DNA Transposable Elements; 0/DNA, Recombinant; EC 2.7.1.-/Phosphoenolpyruvate Sugar Phosphotransferase System; EC 2.7.1.-/phosphoenolpyruvate-mannose phosphotransferase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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