Document Detail


Monitoring enzyme expression of a branched respiratory chain of corynebacterium glutamicum using an EGFP reporter gene.
MedLine Citation:
PMID:  21643696     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
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.
Authors:
Tomoichirou Kusumoto; Makoto Aoyagi; Hideo Iwai; Yoshiki Kabashima; Junshi Sakamoto
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-6-4
Journal Detail:
Title:  Journal of bioenergetics and biomembranes     Volume:  -     ISSN:  1573-6881     ISO Abbreviation:  -     Publication Date:  2011 Jun 
Date Detail:
Created Date:  2011-6-6     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7701859     Medline TA:  J Bioenerg Biomembr     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
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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