Compare a wild type Saccaromyces cerevisise strain V5 strain (=V), with a mutant strain where glucose transport has been reduced by at least 30% in the strain V5.TM6*P (=T), mutant is lacking 7 hexose transporters HXT1, HXT2, HXT3, HXT4, HXT5, HXT6, HXT7, the plasma membrane contains a HXT1-HXT7 fusion protein. This construct is a combination of a low and high affinity glucose transporters. The glucose concentration in the culture media of each array is the parameter that can be how the arrays are sorted. Yeast cells grown in bioreactors monitoring, pH (=5) temp (=30ºC) and pO2>50% as well as O2 and CO2 in outlet gas. Growth media is YNB-CSM starting at 5% glucose. No detectable or small levels of glycerol (0g/g-starting glucose concentration) and ethanol (<2.5g/g-starting glucose concentration) after glucose depletion in the V5.TM6*P supporting the claim that V5.TM6*P grows fully respiratory on glucose. Result 1: V5 to V5.TM6*P RNA transition induce genes typical for respiratory growth. Result 2: There is a glucose dependence in the array signal of 20 genes (e.g HXT1, RGS2, ADH7, CHA4 and GLK1) in wild type V5 strain is not seen in the V5.TM6*P strain, this is an evidence for that glucose sensing occurs inside the yeast cell. That is, an intact glucose plasma membrane transport system is required for responding to a change in glucose concentration in the range 40 to 5g/L glucose. Last three letters of title is glucose concentration in media, in units g/L if divided by 10 (e.g. T1_MUTANT_13_YA0403#22_365, the residual glucose concentration of this sample is 36.5g/L).
Treatment protocol
Transformants were selected by growth on glucose. The resulting ura3 strain is referred to as V5.TM6*. The auxotrophic V5 wild type and V5.TM6* used in this study were made prototrophic by integration of URA3 [3] and are referred to as the V5 wild-type strain and V5.TM6*P, respectively. The growth media in the bioreactor was 5 × defined minimal medium [4], stirring was 1000rpm; Temp 30C; Aeriation 0.5 vvm; pH 5.
Extracted molecule
total RNA
Extraction protocol
Wistar Institute extract protocol
Label
P33
Label protocol
Wistar Institute label protocol
Hybridization protocol
Wistar Institute Hybridization protocol
Scan protocol
Wistar Institute scan protocol
Description
na
Data processing
Wistar Institute nMD: normalized Median Density method
