Abstract
Combining high-throughput growth physiology and global gene expression data analysis is of significant value for integrating metabolism and genomics. We compared global gene expression using 500 ng of total RNA from Escherichia coli cultures grown in rich or defined minimal media in a miniaturized 50-μl bioreactor. The microbioreactor was fabricated out of poly(dimethylsiloxane) (PDMS) and glass and equipped to provide on-line, optical measurements. cDNA labeling for microarray hybridizations was performed with the GeniconRLS system. From these experiments, we found that the expression of 232 genes increased significantly in cells grown in minimum medium, including genes involved in amino acid biosynthesis and central metabolism. The expression of 275 genes was significantly elevated in cells grown in rich medium, including genes involved in the translational and motility apparatuses. In general, these changes in gene expression levels were similar to those observed in 1,000-fold larger cultures. The increasing rate at which complete genomic sequences of microorganisms are becoming available offers an unprecedented opportunity for investigating these organisms. Our results from microscale cultures using just 500 ng of total RNA indicate that high-throughput integration of growth physiology and genomics will be possible with novel biochemical platforms and improved detection technologies.
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Acknowledgements
We gratefully acknowledge the DuPont-MIT Alliance (DMA) for funding. The authors are also grateful to Dr. Jefferson Parker for his help with gene annotation, Dr. Phil Lessard for continuous, stimulating discussions and for reviewing the manuscript, and Prof. Jacqueline Piret for critically reviewing the manuscript.
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Boccazzi, P., Zanzotto, A., Szita, N. et al. Gene expression analysis of Escherichia coli grown in miniaturized bioreactor platforms for high-throughput analysis of growth and genomic data. Appl Microbiol Biotechnol 68, 518–532 (2005). https://doi.org/10.1007/s00253-005-1966-6
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DOI: https://doi.org/10.1007/s00253-005-1966-6