Development of real-time PCR primer and probe sets for detecting degenerated and non-degenerated forms of the butanol-producing bacterium Clostridium acetobutylicum ATCC 824.
Degeneration is one of the limiting factors in butanol fermentation, and it must be monitored and prevented for stable butanol production. In Clostridium acetobutylicum ATCC 824, the most well-known butanol-producing microorganism, degeneration is caused by the loss of the pSOL1 plasmid that carries essential genes involved in solvent production. In this study, we designed two specific primer and probe sets for real-time qPCR (RT-qPCR) detection of C. acetobutylicum ATCC 824 (the C. aceto set) and pSOL1-possessing C. acetobutylicum ATCC 824 (the DGS set). Specific primer and probe sets were designed on the basis of the 16S rDNA sequence and pSOL1 sequence. The number of degenerated C. acetobutylicum could be quantified by subtracting the number of C. acetobutylicum ATCC 824 containing pSOL1 from the total number of C. acetobutylicum ATCC 824. The primer and probe sets permitted the specific detection and quantification of degenerated C. acetobutylicum and total butanol-producing C. acetobutylicum by RT-qPCR.
Lee
Sun-Mi
SM
Center for Environmental Technology Research, Korea Institute of Science and Technology, Seoul, Republic of Korea.
Cho
Min Ok
MO
Um
Youngsoon
Y
Sang
Byoung-In
BI
D000440:Butanols / Q000378:metabolism*; D046969:Clostridium acetobutylicum / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics*; D005285:Fermentation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D000440:Butanols; D017931:DNA Primers; D015342:DNA Probes