U.S. Department of Energy Office of Biological and Environmental Research

BER Research Highlights


Promoter Set for Heterologous Gene Expression in Clostridium thermocellum
Published: March 30, 2015
Posted: April 20, 2015

For successful fermentation of biofuels and bioproducts from biomass, using microorganisms for which fewer genetic tools have been developed might be the most effective approach. To date, most metabolic engineering work in Clostridium thermocellum has focused on gene deletion, but many metabolic engineering strategies require well controlled heterologous gene expression, which requires a collection of well characterized and understood promoters. Researchers from the Department of Energy’s BioEnergy Science Center sought to identify new promoters for predictable gene expression in C. thermocellum. For this work, 17 different C. thermocellum promoters were tested with two different reporter genes (LacZ and AdhB) to ensure the activity of the target promoter was not gene-specific. Putative promoters were chosen by analyses of published C. thermocellum gene expression datasets. Promoter activity in both C. thermocellum and Escherichia coli were testedbecauseideally a promoter would not be strongly expressed in E. coli to avoid toxicity problems during cloning. Several useful promoters were identified (eno, cbp, cbp_2, 815, 966, 2638, and 2926), which showed high expression and high enzymatic activity of both reporter genes in C. thermocellum. Other promoters were not useful, showing no heterologous gene activity or negatively impacting plasmid stability. These results provide several new good promoters for C. thermocellum. This improved understanding of promoter function will enhance efforts to express heterologous genes important for improved biofuel production in C. thermocellum.

Reference: Olson, D. G., M. Maloney, A. A. Lanahan, S. Hon, L. J. Hauser, and L. R. Lynd. 2015. “Identifying Promoters for Gene Expression in Clostridium thermocellum,” Metabolic Engineering Communications, DOI: 10.1016/j.meteno.2015.03.002. (Reference link)

Contact: Kent Peters, SC-23.2, (301) 903-5549
Topic Areas:

  • Research Area: Genomic Analysis and Systems Biology
  • Research Area: Microbes and Communities
  • Research Area: Sustainable Biofuels and Bioproducts
  • Research Area: DOE Bioenergy Research Centers (BRC)
  • Research Area: Biosystems Design

Division: SC-23.2 Biological Systems Science Division, BER

 

BER supports basic research and scientific user facilities to advance DOE missions in energy and environment. More about BER

Recent Highlights

Aug 24, 2019
New Approach for Studying How Microbes Influence Their Environment
A diverse group of scientists suggests a common framework and targeting of known microbial processes [more...]

Aug 08, 2019
Nutrient-Hungry Peatland Microbes Reduce Carbon Loss Under Warmer Conditions
Enzyme production in peatlands reduces carbon lost to respiration under future high temperatures. [more...]

Aug 05, 2019
Amazon Forest Response to CO2 Fertilization Dependent on Plant Phosphorus Acquisition
AmazonFACE Model Intercomparison. The Science Plant growth is dependent on the availabi [more...]

Jul 29, 2019
A Slippery Slope: Soil Carbon Destabilization
Carbon gain or loss depends on the balance between competing biological, chemical, and physical reac [more...]

Jul 15, 2019
Field Evaluation of Gas Analyzers for Measuring Ecosystem Fluxes
How gas analyzer type and correction method impact measured fluxes. The Science A side- [more...]

List all highlights (possible long download time)