TY - JOUR
T1 - Visualization of differential gene expression by improved cyan fluorescent protein and yellow fluorescent protein production in Bacillus subtilis
AU - Veening, Jan Willem
AU - Smits, Wiep Klaas
AU - Hamoen, Leendert W.
AU - Jongbloed, Jan D.H.
AU - Kuipers, Oscar P.
PY - 2004/11/1
Y1 - 2004/11/1
N2 - The distinguishable cyan and yellow fluorescent proteins (CFP and YFP) enable the simultaneous in vivo visualization of different promoter activities. Here, we report new cloning vectors for the construction of cfp and yfp fusions in Bacillus subtilis. By extending the N-terminal portions of previously described CFP and YFP variants, 20- to 70-fold-improved fluorescent-protein production was achieved. Probably, the addition of sequences encoding the first eight amino acids of the N-terminal part of ComGA of B. subtilis overcomes the slow translation initiation that is provoked by the eukaryotic codon bias present in the original cfp and yfp genes. Using these new vectors, we demonstrate that, within an isogenic population of sporulating B. subtilis cells, expression of the abrB and spoIIA genes is distinct in individual cells.
AB - The distinguishable cyan and yellow fluorescent proteins (CFP and YFP) enable the simultaneous in vivo visualization of different promoter activities. Here, we report new cloning vectors for the construction of cfp and yfp fusions in Bacillus subtilis. By extending the N-terminal portions of previously described CFP and YFP variants, 20- to 70-fold-improved fluorescent-protein production was achieved. Probably, the addition of sequences encoding the first eight amino acids of the N-terminal part of ComGA of B. subtilis overcomes the slow translation initiation that is provoked by the eukaryotic codon bias present in the original cfp and yfp genes. Using these new vectors, we demonstrate that, within an isogenic population of sporulating B. subtilis cells, expression of the abrB and spoIIA genes is distinct in individual cells.
UR - http://www.scopus.com/inward/record.url?scp=8744302067&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=8744302067&partnerID=8YFLogxK
U2 - 10.1128/AEM.70.11.6809-6815.2004
DO - 10.1128/AEM.70.11.6809-6815.2004
M3 - Article
C2 - 15528548
AN - SCOPUS:8744302067
SN - 0099-2240
VL - 70
SP - 6809
EP - 6815
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
IS - 11
ER -