Paul Magnano:Week 3 Individual Journal: Difference between revisions
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==Outline== | ==Outline== | ||
===Experiment=== | |||
*sacchromyces cerevisiae was grown in continuous culture, with varying ammonia input concentrations, while ammonia assiilation rates remained constant | |||
*ammonia is a preffered nitrogen source for s. cerevisiae, because it results in faster growth than other sources | |||
*in this study, cultures were grown with same flux (relationship b/w external ammonia concentration and rte of assimilation) but different inflow ammonia concentrations | |||
*Physiological parameters: | |||
**S. cerevisiae SU32 grown in continuous cultures with ammonia inflow concentrations ranging from 29 to 118mM | |||
**fixed glucose concentration of 100mM | |||
*Fig 1A | |||
**increase of ammonia concentration from 29 to 61mM resulted in increase of biomass from 4.9 to 8.2 g/liter | |||
**ammonia concentration above 61mM biomass remained at 8.2 g/liter | |||
**over trhe entire range of ammonia concentrations the ammonia flux into biomass was 1.1 mmol/gh | |||
*Fig 1B | |||
**input ammonia concentration above 44mM resulted in (c02 production/02 consumption) remained constant | |||
**with ammonia limitation (ammonia input below 44mM) c02 production and 02 consumption values differed | |||
**no changes in the residual glucose concentration were seen | |||
**no significant changes in carbon metabolism occured when the culture was switched from ammonia excess to ammonia limitation | |||
*Fig 1C | |||
**ammonia within the cells reacts with ketoglutarate to produce glutamate which is converted to glutamine | |||
**ketoglutarate concentration decreased from 10 to 5 umol/g when the ammonia concentration changed from limited to excess | |||
**glutamate concentration increased within the cell, linearly | |||
**increaing ammonia concentrations caused the concentrations of glutamate and glutamine within the cell to increase | |||
===Northern Analyses=== | |||
*RNA analyses were done to see if RNA levels of nitrogen regulating genes changed with increased ammonia concentrations | |||
*genes GDH1, GDH2,GLN1 were determined to study responses to ammonia levels | |||
*Fig 2 | |||
**increasing ammonia levels resulted in GDH1 levels staying the same | |||
**ammonia concentrations of 29-44mM there was no GDH2, but with increase to 61mM GDH2 level increased | |||
**maximum GLN1 expression was observed at 61mM | |||
** overall it was seen that ammonia concentration both repressed GDH1 and caused the expression of GDH2 | |||
==Ten New Terms== | ==Ten New Terms== |
Revision as of 20:10, 30 January 2013
Outline
Experiment
- sacchromyces cerevisiae was grown in continuous culture, with varying ammonia input concentrations, while ammonia assiilation rates remained constant
- ammonia is a preffered nitrogen source for s. cerevisiae, because it results in faster growth than other sources
- in this study, cultures were grown with same flux (relationship b/w external ammonia concentration and rte of assimilation) but different inflow ammonia concentrations
- Physiological parameters:
- S. cerevisiae SU32 grown in continuous cultures with ammonia inflow concentrations ranging from 29 to 118mM
- fixed glucose concentration of 100mM
- Fig 1A
- increase of ammonia concentration from 29 to 61mM resulted in increase of biomass from 4.9 to 8.2 g/liter
- ammonia concentration above 61mM biomass remained at 8.2 g/liter
- over trhe entire range of ammonia concentrations the ammonia flux into biomass was 1.1 mmol/gh
- Fig 1B
- input ammonia concentration above 44mM resulted in (c02 production/02 consumption) remained constant
- with ammonia limitation (ammonia input below 44mM) c02 production and 02 consumption values differed
- no changes in the residual glucose concentration were seen
- no significant changes in carbon metabolism occured when the culture was switched from ammonia excess to ammonia limitation
- Fig 1C
- ammonia within the cells reacts with ketoglutarate to produce glutamate which is converted to glutamine
- ketoglutarate concentration decreased from 10 to 5 umol/g when the ammonia concentration changed from limited to excess
- glutamate concentration increased within the cell, linearly
- increaing ammonia concentrations caused the concentrations of glutamate and glutamine within the cell to increase
Northern Analyses
- RNA analyses were done to see if RNA levels of nitrogen regulating genes changed with increased ammonia concentrations
- genes GDH1, GDH2,GLN1 were determined to study responses to ammonia levels
- Fig 2
- increasing ammonia levels resulted in GDH1 levels staying the same
- ammonia concentrations of 29-44mM there was no GDH2, but with increase to 61mM GDH2 level increased
- maximum GLN1 expression was observed at 61mM
- overall it was seen that ammonia concentration both repressed GDH1 and caused the expression of GDH2