Colony-live --a high-throughput method for measuring microbial colony growth kinetics-- reveals diverse growth effects of gene knockouts in Escherichia coli
| Autor: | Takeyuki Tamura, Ai Muto, Yuichirou Tanaka, Rikiya Takeuchi, Barry L. Wanner, Toru Nakayashiki, Hirotada Mori |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Genetics
Microbial Microbiology (medical) Single-gene knockout Lag time of growth (LTG) Growth kinetics High-throughput Computational biology Biology medicine.disease_cause Microbiology Phenotype screening Gene Knockout Techniques Optical imaging Saturation point growth (SPG) High-Throughput Screening Assays Escherichia coli medicine Gene Throughput (business) Gene knockout Genetics Bacteriological Techniques Methodology Article Optical Imaging Keio collection Maximum growth rate (MGR) |
| Zdroj: | BMC Microbiology |
| ISSN: | 1471-2180 |
| Popis: | Background Precise quantitative growth measurements and detection of small growth changes in high-throughput manner is essential for fundamental studies of bacterial cell. However, an inherent tradeoff for measurement quality in high-throughput methods sacrifices some measurement quality. A key challenge has been how to enhance measurement quality without sacrificing throughput. Results We developed a new high-throughput measurement system, termed Colony-live. Here we show that Colony-live provides accurate measurement of three growth values (lag time of growth (LTG), maximum growth rate (MGR), and saturation point growth (SPG)) by visualizing colony growth over time. By using a new normalization method for colony growth, Colony-live gives more precise and accurate growth values than the conventional method. We demonstrated the utility of Colony-live by measuring growth values for the entire Keio collection of Escherichia coli single-gene knockout mutants. By using Colony-live, we were able to identify subtle growth defects of single-gene knockout mutants that were undetectable by the conventional method quantified by fixed time-point camera imaging. Further, Colony-live can reveal genes that influence the length of the lag-phase and the saturation point of growth. Conclusions Measurement quality is critical to achieving the resolution required to identify unique phenotypes among a diverse range of phenotypes. Sharing high-quality genome-wide datasets should benefit many researchers who are interested in specific gene functions or the architecture of cellular systems. Our Colony-live system provides a new powerful tool to accelerate accumulation of knowledge of microbial growth phenotypes. |
| Databáze: | OpenAIRE |
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