Monitoring enzyme expression of a branched respiratory chain of <Emphasis Type="Italic">corynebacterium glutamicum</Emphasis> using an EGFP reporter gene |
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Authors: | Tomoichirou?Kusumoto Makoto?Aoyagi Hideo?Iwai Yoshiki?Kabashima Email author" target="_blank">Junshi?SakamotoEmail author |
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Institution: | Department of Bioscience and Bioinformatics, Kyushu Institute of Technology, 680-4 Kawazu, Iizuka, Fukuoka, 820-8502, Japan. tomo@bio.kyutech.ac.jp |
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Abstract: | To investigate the expressional control of branched respiratory chain complexes of the amino-acid producing bacterium Corynebacterium glutamicum according to growth conditions, the expression indexes of the ndh, sdh, qcrCAB, ctaCF, ctaD, ctaE, and cydAB genes were estimated under aerobic and microaerobic, and carbon-rich and -poor conditions. The promoter region of each target
gene was cloned upstream of the EGFP gene on expression vector pVK6, and the nine reporter constructs were transformed into
C. glutamicum ssp. lactofermentum. The cytochrome content of cellular membranes obtained from each growth phase closely corresponded to the expression indexes
based on EGFP fluorescence and cell density, indicating that this rapid and convenient method is suitable for analyzing the
expression levels of respiratory chain complexes. Using this method, we demonstrated that a reciprocal change in the expression
levels of cytochrome bd-type and aa
3-type oxidases occurs when C. glutamicum cells are held in stationary phase for extended periods. |
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