Improved lux reporters for use in Staphylococcus aureus.
The use of luxABCDE (lux) offers certain advantages over other reporters, such as: lacZ and xylE. It is real time and its signal generation is produced without the requirement for any additional substrates. In some bacteria such as Staphylococcus spp, light production by luciferase is restricted because of a limited availability of endogenous substrates such as fatty acid aldehyde. We describe the construction of promoterless-lux cloning vectors, pGYlux and pAmilux. S. aureus carrying B. subtilis xyl/tetO promoter fused to the lux genes of pGYlux gave up to a 2.5-fold enhancement of luminescence over S. aureus carrying the xyl/tetO promoter fused to lux genes of the previously published parent vector pAL2. Furthermore, pAmilux showed a 6-fold enhancement of lux expression when compared to pGYlux in S. aureus. This was achieved by cloning the constitutive ami promoter upstream of the luxCDE genes to increase endogenous fatty acid aldehyde production while maintaining its reporter functionality by fusing promoters to the luxAB genes.
Mesak
Lili Rosana
LR
Department of Microbiology and Immunology, Life Sciences Institute, University of British Columbia, 2350 Health Science Mall, Vancouver, Canada BC V6T 1Z3. mesak@interchange.ubc.ca
Yim
Grace
G
Davies
Julian
J
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D008156:Luciferases; D049449:Luminescence; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D013211:Staphylococcus aureus / Q000235:genetics*
D017931:DNA Primers; D008156:Luciferases