Differential gene expression profiles and real-time measurements of growth parameters in Saccharomyces cerevisiae grown in microliter-scale bioreactors equipped with internal stirring.
Combining real-time growth kinetics measurements with global gene expression analysis of microbial cultures is of significant value for high-throughput biological research. We have performed differential gene expression analysis in the eukaryotic model Saccharomyces cerevisiae grown in galactose and glucose media in 150 muL bioreactors equipped with sensors for in situ and real-time measurements of optical density (OD), pH, and dissolved oxygen (DO). The microbioreactors were fabricated from poly(dimethylsiloxane) (PDMS) and poly(methyl methacrylate) (PMMA) and equipped with internal magnetic ministirrers and evaporation compensation by water replacement. In galactose-grown cells, the core genes of the GAL operon GAL2, GAL1, GAL7, and GAL10 were upregulated at least 100-fold relative to glucose-grown cells. These differential gene expression levels were similar to those observed in large-scale culture vessels. The increasing rate at which complete genomic sequences of microorganisms are becoming available offers an unprecedented opportunity for comparative investigations of these organisms. Our results from S. cerevisiae cultures grown in instrumented microbioreactors show that it is possible to integrate high-throughput studies of growth physiology with global gene expression analysis of microorganisms.
Boccazzi
Paolo
P
Department of Biology and Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Zhang
Zhiyu
Z
Kurosawa
Kazuhiko
K
Szita
Nicolas
N
Bhattacharya
Sanchita
S
Jensen
Klavs F
KF
Sinskey
Anthony J
AJ
D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis / Q000379:methods; D005690:Galactose / Q000737:chemistry / Q000494:pharmacology; D020869:Gene Expression Profiling; D005947:Glucose / Q000737:chemistry / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D010100:Oxygen / Q000737:chemistry; D012313:RNA / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D013997:Time Factors
D012313:RNA; D005947:Glucose; D010100:Oxygen; D005690:Galactose