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Published on: October 2, 2012
Discovery of a highly active condition-responsive promoter for constructing a C1 gas-fermenting platform based on
Byeongchan Kang1, Ji-Yeon Kim1, Soyoung Oh1
1Department of Environment and Energy Engineering, Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Jeonnam-Gwangju 61005, the Republic of Korea; Research Center for Innovative Energy and Carbon Optimized Synthesis for Chemicals (inn-ECOSysChem), Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Jeonnam-Gwangju 61005, the Republic of Korea.
Abstract:
Integrating differential RNA-seq profiling with de novo motif discovery in Eubacterium callanderi KIST612 identified distinct sequence features associated with substrate-responsive transcription. Specifically, MOTIF4 and MOTIF17 showed significant correlations with glucose- and C1 gas (CO)-responsive gene expression, respectively. Based on these motif profiles, a candidate CO-responsive promoter, CI04, was selected and experimentally validated in vivo using a β-glucuronidase (GUS) reporter assay. CI04 exhibited specific CO-responsive activity, exceeding 500 mU mg-1 under CO conditions. Under CO conditions, CI04 activity was approximately 10-fold higher than under glucose conditions, 59-fold higher than that of the constitutive (CT) promoter, and approximately 2.5-fold higher under CO conditions than that of the H2 promoter previously used for overexpression in E. callanderi KIST612.
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