earticle

영어

Clostridium beijerinckii is a gram-positive, spore-forming, obligate anaerobic bacterium, which is able to produce solvents acetone, butanol and ethanol by using various sugars. A gene expression reporter system (pBEst-MTL) for C. beijerinckii NCIMB 8052 was developed by using esterase gene from Bacillus subtilis as a reporter gene. In order to test the reporter system, promoters of two key metabolic pathway genes, ptb (coding for phosphotransbutyrylase), thl
(coding for thiolase), were cloned upstream of the reporter gene in pBEst-MTL in order to construct plasmids pBEst-MBP and pBEst-MBT, respectively. Detection of esterase activity in time course studies performed with strains NCIMB 8052 [pBEst-MBP], NCIMB 8052 [pBEst-MBP] demonstrated that the reporter gene produced a functional esterase in C. beijerinckii. In addition, time course studies revealed differences in esterase specific activity profiles of strains
NCIMB 8052 [pBEst-MBP], NCIMB 8052 [pBEst-MBP], suggesting that the reporter system developed in this study is able to effectively distinguish between different promoters.