Cells were grown to log phase at 23°C before being shifted to 37°C at time zero.
Aliquots of 500 µl from the cultures of S. mutans ATCC 25175 grown to log phase were added to these tubes.
To evaluate growth in CM, GBS was initially grown to log phase (OD600 0.3) in THB.
Briefly, yeast strains expressing TAP-tagged subunits were grown to log phase and whole cell extracts were prepared.
Yeast cells were grown to log phase and fixed with 3.7% formaldehyde for 15 min at 30°C.
Briefly, CC-125 cells grown to log phase were collected and the cell density was determined with a hemacytometer.
This culture was then back-diluted to OD600 of 0.1 and then grown to log phase (OD600 of 0.6±0.1).
Briefly, 32 liters of CC-125 cells grown to log phase in TAP medium were collected and resuspended in 4 liters of 10 mM HEPES, pH 7.2.
Bacteria were then re-inoculated in CM containing 1% pullulan and grown to log phase (OD600 0.4) to allow the expression of pullulanases on bacterial surface.
At hour 12, into the 5ml aliquot of Tube 1 and Tube 4 (107/ml), approximately 50/ml transductant cells (BNT1080 orBNT1081) grown to log phase were added.
The transformed cells were grown to log phase (OD600 = 0.6 to 1.0) and IPTG was added to a final concentration of 0.4 mM.
