Construction and use of GFP reporter vectors for analysis of cell-type-specific gene expression in Nostoc punctiforme

Journal of Microbiological Methods
Claudia ArguetaMichael L Summers

Abstract

Two transcriptional reporter shuttle vectors were constructed for the filamentous cyanobacterium Nostoc punctiforme using the green fluorescence protein (GFP) reporter. Both the ampicillin- and kanamycin-resistant versions of the plasmid allow promoters to be directionally cloned into a multiple cloning site preceding a promoterless gfp gene using an Escherichia coli host. The ability of the self-replicating shuttle plasmids to report cell-type-specific gene expression in N. punctiforme was tested by cloning promoters expressed in normal vegetative cells, nitrogen-fixing heterocysts and spore-like akinetes. A P(psaC) reporter gene fusion was expressed in vegetative cells and not in heterocysts, whereas GFP driven from P(hetR) was found highly expressed in heterocysts. GFP expression driven by the promoter for the N. punctiforme homologue of the akinete-specific gene avaK was expressed in developing akinetes. Decreased expression of GFP from the P(psaC) reporter in hormogonia was also observed. The results demonstrate the utility of these GFP vectors to study cell-type-specific gene expression in differentiating filamentous cyanobacteria.

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Citations

May 21, 2005·Archives of Microbiology·Claudia Argueta, Michael L Summers
May 29, 2012·Origins of Life and Evolution of the Biosphere : the Journal of the International Society for the Study of the Origin of Life·Daniela Billi
Jun 7, 2011·Journal of Bacteriology·Paulo Oliveira, Peter Lindblad
Mar 17, 2009·BMC Microbiology·Marie HolmqvistPeter Lindblad
Jul 31, 2014·Nucleic Acids Research·Arnaud TatonJames W Golden
Dec 16, 2010·FEBS Letters·Yoon-Jung MoonYoung-Ho Chung
Aug 21, 2015·Applied Microbiology and Biotechnology·L HudekM L Ackland
Jun 20, 2006·Molecular Microbiology·Claudia ArguetaMichael L Summers
Oct 1, 2011·Environmental Microbiology Reports·Anton LiaimerElke Dittmann
Dec 2, 2014·Applied Microbiology and Biotechnology·Kangming ChenRuanbao Zhou
Feb 14, 2013·PloS One·Saakshi JalaliVinod Scaria
Oct 12, 2010·Journal of Child Psychology and Psychiatry, and Allied Disciplines·Jennifer Barrett, Alison S Fleming

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