Gene expression and survival changes in Saccharomyces cerevisiae during suspension culture.

This study explores the connection between changes in gene expression and the genes that determine strain survival during suspension culture, using the model eukaryotic organism, Saccharomyces cerevisiae. The Saccharomyces cerevisiae homozygous diploid deletion pool, and the BY4743 parental strain were grown for 18 hours in a rotating wall vessel, a suspension culture device optimized to minimize the delivered shear. In addition to the reduced shear conditions, the rotating wall vessels were also placed in a static position or in a shaker in order to change the amount of shear stress on the cells. Keywords: shear stress, time course The parental and Homozygous diploid pool were each pipetted into 6 RWVs per strain. For this study, the type of RWV used was the 10 mL HARV, which is designed to provide continuous O2-CO2 gas exchange across the membrane. Residual air was removed through the syringe port after loading. All RWVs were then transferred to a 30 °CC incubator. Within the incubator, two RWVs per strain were placed on rotators set at 30 rpm, two RWVs were placed in a gyrorotatory shaker incubator agitated at 90 rpm coincidental with the gravity vector, and the remaining two RWVs were allowed to remain static in a horizontal position. All RWVs were grown for 18 hours before harvet. RNA was extracted from the BY4743 samples, DNA from the deletion pool and used to generate probes for gene expression or DNA Tag3 arrays.