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      The application of Tet repressor in prokaryotic gene regulation and expression

      review-article
      1 , 2 , * , 2
      Microbial Biotechnology
      Blackwell Publishing Ltd

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          Summary

          Inducible gene expression based upon Tet repressor ( tet regulation) is a broadly applied tool in molecular genetics. In its original environment, Tet repressor (TetR) negatively controls tetracycline (tc) resistance in bacteria. In the presence of tc, TetR is induced and detaches from its cognate DNA sequence tetO, so that a tc antiporter protein is expressed. In this article, we provide a comprehensive overview about tet regulation in bacteria and illustrate the parameters of different regulatory architectures. While some of these set‐ups rely on natural tet‐control regions like those found on transposon Tn 10, highly efficient variations of this system have recently been adapted to different Gram‐negative and Gram‐positive bacteria. Novel tet‐controllable artificial or hybrid promoters were employed for target gene expression. They are controlled by regulators expressed at different levels either in a constitutive or in an autoregulated manner. The resulting tetsystems have been used for various purposes. We discuss integrative elements vested with tc‐sensitive promoters, as well as tet regulation in Gram‐negative and Gram‐positive bacteria for analytical purposes and for protein overproduction. Also the use of TetR as an in vivo biosensor for tetracyclines or as a regulatory device in synthetic biology constructs is outlined. Technical specifications underlying different regulatory set‐ups are highlighted, and finally recent developments concerning variations of TetR are presented, which may expand the use of prokaryotic tet systems in the future.

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          Most cited references105

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          Cohesins: chromosomal proteins that prevent premature separation of sister chromatids.

          Cohesion between sister chromatids opposes the splitting force exerted by microtubules, and loss of this cohesion is responsible for the subsequent separation of sister chromatids during anaphase. We describe three chromosmal proteins that prevent premature separation of sister chromatids in yeast. Two, Smc1p and Smc3p, are members of the SMC family, which are putative ATPases with coiled-coil domains. A third protein, which we call Scc1p, binds to chromosomes during S phase, dissociates from them at the metaphase-to-anaphase transition, and is degraded by the anaphase promoting complex. Association of Scc1p with chromatin depends on Smc1p. Proteins homologous to Scc1p exist in a variety of eukaryotic organisms including humans. A common cohesion apparatus might be used by all eukaryotic cells during both mitosis and meiosis.
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            Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems.

            Kay Terpe (2006)
            During the proteomics period, the growth in the use of recombinant proteins has increased greatly in the recent years. Bacterial systems remain most attractive due to low cost, high productivity, and rapid use. However, the rational choice of the adequate promoter system and host for a specific protein of interest remains difficult. This review gives an overview of the most commonly used systems: As hosts, Bacillus brevis, Bacillus megaterium, Bacillus subtilis, Caulobacter crescentus, other strains, and, most importantly, Escherichia coli BL21 and E. coli K12 and their derivatives are presented. On the promoter side, the main features of the l-arabinose inducible araBAD promoter (PBAD), the lac promoter, the l-rhamnose inducible rhaP BAD promoter, the T7 RNA polymerase promoter, the trc and tac promoter, the lambda phage promoter p L , and the anhydrotetracycline-inducible tetA promoter/operator are summarized.
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              Allelic replacement in Staphylococcus aureus with inducible counter-selection.

              A method for rapid selection of allelic replacement mutations in the chromosome of Staphylococcus aureus is described. Plasmid pKOR1, an Escherichia coli/S. aureus shuttle vector, permits rapid cloning via lambda recombination and ccdB selection. Plasmid transformation of staphylococci and growth at 43 degrees C, a non-permissive condition for pKOR1 replication, selects for homologous recombination and pKOR1 integration into the bacterial chromosome. Anhydrotetracycline-mediated induction of pKOR1-encoded secY antisense transcripts via the Pxyl/tetO promoter, a condition that is not compatible with staphylococcal growth, selects for chromosomal excision and loss of plasmid. Using this strategy, allelic replacements in S. aureus rocA were generated at frequencies that obviated the need for antibiotic marker selection.
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                Author and article information

                Journal
                Microb Biotechnol
                Microb Biotechnol
                MBT
                Microbial Biotechnology
                Blackwell Publishing Ltd (Oxford, UK )
                1751-7915
                1751-7915
                January 2008
                24 August 2007
                : 1
                : 1
                : 2-16
                Affiliations
                [1 ]Lehrbereich Mikrobielle Genetik, Eberhard Karls Universität Tübingen, Waldhäuserstr. 70/8, 72076 Tübingen, Germany.
                [2 ]Lehrstuhl für Mikrobiologie, Institut für Biologie, Friedrich‐Alexander‐Universität Erlangen‐Nürnberg, Staudtstrasse 5, 91058 Erlangen, Germany.
                Author notes
                *E‐mail ralph.bertram@ 123456uni‐tuebingen.de ; Tel. (+49) 7071 29 78855; Fax (+49) 7071 29 5937.
                Article
                10.1111/j.1751-7915.2007.00001.x
                3864427
                21261817
                fa9432c1-93b1-4ad1-afc1-5f5c1fefa38d
                Copyright © 2007 The Authors. Journal compilation © 2007 Society for Applied Microbiology and Blackwell Publishing Ltd.
                History
                : 24 April 2007
                : 29 May 2007
                : 30 May 2007
                Categories
                Review

                Biotechnology
                Biotechnology

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