Test of anaerobic fluorescent protein as reporter of gene expression in Clostridium sp. AWRP

Abstract

Acetogenic bacteria are important industrial microorganisms that produce various chemicals and fuels. Effective genetic tools are still needed for improved chemical production through metabolic engineering. Anaerobic fluorescent protein evoglow as reporter gene is used to monitor gene expression in some anaerobes. In this study, we tested evoglow expression under phosphotransbutyrylase promoter (Pptb) of C. acetobutylicum in Clostridium sp. AWRP, a newly isolated acetogenic species. The recombinant expression vectors were introduced into Clostridium sp. AWRP via conjugation. The expression was analyzed by fluorescent microscopy. Here, we showed that the fluorescence intensity was strong in Clostridium sp. AWRP codon- optimized evoglow gene. These results suggest that the anaerobic fluorescent protein evoglow can be used for effective monitoring gene expression in Clostridium sp. AWRP.glow as reporter gene is used to monitor gene expression in some anaerobes. In this study, we tested evoglow expression under phosphotransbutyrylase promoter (Pptb) of C. acetobutylicum in Clostridium sp. AWRP, a newly isolated acetogenic species. The recombinant expression vectors were introduced into Clostridium sp. AWRP via conjugation. The expression was analyzed by fluorescent microscopy. Here, we showed that the fluorescence intensity was strong in Clostridium sp. AWRP codon- optimized evoglow gene. These results suggest that the anaerobic fluorescent protein evoglow can be used for effective monitoring gene expression in Clostridium sp. AWRP.