Tight control of transcription in Toxoplasma gondii using an alternative tet repressor

International Journal for Parasitology
Nicole F J van PoppelDick Schaap

Abstract

Fusion of yellow fluorescent protein (YFP) to the N-terminus of the Escherichia coli Tn10 tet repressor (TetR) created a functional YFP-TetR repressor with the capacity of 88-fold repression of transcription when expressed in Toxoplasma gondii. As a test promoter we used the T. gondii ribosomal protein RPS13 promoter for which we provide experimental evidence of having a single major transcriptional start site, a condition favourable to the design of inducible expression systems. Integration of four tet operator (tetO) elements, 23-43 bp upstream of the RPS13 transcriptional start site, resulted in maximal repression of transcription (88-fold). Moreover, integration of these four tetO elements reduced the promoter activity only 20% in comparison with the wildtype promoter. Regulation was six-fold higher compared with an inducible expression system employing wildtype TetR. Importantly, only 0.1 microg/ml tetracycline was required for maximal induction demonstrating a higher affinity of tetracycline for YFP-TetR than for wildtype TetR which required 1 microg/ml tetracycline for maximal induction. The use of 0.1 microg/ml tetracycline allows prolonged continuous culturing of T. gondii for which levels of 1 microg/ml tetracycline a...Continue Reading

References

Mar 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·C Gatz, P H Quail
Jan 1, 1997·Molecular and Biochemical Parasitology·L HamannE Tannich
Jan 1, 1997·Molecular and Biochemical Parasitology·G RamakrishnanW A Petri
Jul 10, 1998·Molecular and Biochemical Parasitology·B StriepenD S Roos
Feb 13, 2001·Molecular and Biochemical Parasitology·S YanK Stuart
Mar 17, 2001·The EMBO Journal·K Ribbeck, D Görlich
Mar 19, 2002·Molecular and Biochemical Parasitology·Noboru InoueJohn E Donelson
Dec 25, 2002·Antimicrobial Agents and Chemotherapy·Marc-Jan GubbelsBoris Striepen
Jan 25, 2005·Molecular and Biochemical Parasitology·Dick SchaapArno N Vermeulen

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Citations

Apr 3, 2007·Applied Microbiology and Biotechnology·Markus MeissnerWilliam J Sullivan
Jan 21, 2014·Experimental Parasitology·Luiz Miguel PereiraAna Patrícia Yatsuda
Nov 3, 2010·Molecular and Biochemical Parasitology·Lauren JammalloMarc-Jan Gubbels
Aug 17, 2014·Molecular and Biochemical Parasitology·Luiz Miguel Pereira, Ana Patrícia Yatsuda
Dec 6, 2014·Journal of Microbiological Methods·Anna K CraterSirinart Ananvoranich
May 18, 2016·Trends in Parasitology·Jin-Lei WangXing-Quan Zhu
Apr 15, 2014·Revista brasileira de parasitologia veterinária = Brazilian journal of veterinary parasitology : Órgão Oficial do Colégio Brasileiro de Parasitologia Veterinária·Luiz Miguel Pereira, Ana Patrícia Yatsuda
Dec 19, 2016·Trends in Parasitology·Edina K Szabo, Constance A M Finney
May 12, 2011·FEBS Letters·Julien Limenitakis, Dominique Soldati-Favre
Jan 6, 2009·The Journal of Eukaryotic Microbiology·Mathieu Gissot, Kami Kim
Aug 10, 2018·Infectious Diseases of Poverty·Cong-Nuan LiuYu-Rong Yang
Sep 27, 2021·Molecular Microbiology·Sylvie BriquetOlivier Silvie

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