Budget Amount *help |
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 2003: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2002: ¥7,700,000 (Direct Cost: ¥7,700,000)
|
Research Abstract |
In order to develop a quantitative model for the main metabolic pathway of Escherichia coli, we cultivated E.coliK12 and W3110, as well as E.coli BW25113 developed in Keio University, and its several single-gene-knockout mutants (zwf^-, pykF^-, gnd^-, ppc^-, pck^-, pgi^- etc.) in batch and continuous modes. We also cultivated several gene knockout mutants (pflA, B, C, D-, pta-, ppc-, adhA- etc.) in microaerobic condition for lactate production. Then we made the metabolic flux analysis based on ^<13>C labeling experiments, measured enzyme activities and the intracellular metabolite concentrations as well as gene expressions using RT-PCR to understand the metabolic regulation quantitatively for the modeling.
|