Article thumbnail

H-NS silences gfp, the green fluorescent protein gene: gfpTCD is a genetically Remastered gfp gene with reduced susceptibility to H-NS-mediated transcription silencing and with enhanced translation

By Colin P. Corcoran, Andrew D. S. Cameron and Charles J. Dorman

Abstract

The bacterial nucleoid-associated protein H-NS, which preferentially targets and silences AT-rich genes, binds the ubiquitous reporter gene gfp and dramatically reduces local transcription. We have redesigned gfp to reduce H-NS-mediated transcription silencing and simultaneously improve translation in vivo without altering the amino acid sequence of the GFP protein. The green fluorescent protein (GFP) from the jellyfish Ae-quorea victoria is widely used as a reporter of transcriptional activity and protein localization in both eukaryotes and pro-karyotes (reviewed in reference 16). The DNA encoding gfp and many of its variants has a high AT content (60%) and strong predicted DNA curvature, both of which are key fea-tures of DNA that is targeted by the histone-like nucleoid structuring protein (H-NS) (10, 12, 14, 15, 22). H-NS is a global gene silencer that represses transcription by condensing chro-mosome and plasmid DNA in many Gram-negative bacteria. For example, H-NS binds and represses 15 % of the Esche

Year: 2010
OAI identifier: oai:CiteSeerX.psu:10.1.1.1024.3421
Provided by: CiteSeerX
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.tara.tcd.ie/bitstre... (external link)
  • http://www.tara.tcd.ie/bitstre... (external link)
  • http://citeseerx.ist.psu.edu/v... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.