Proteins were expressed from Open Biosystems Yeast ORF clones. Clones
were delivered in vials with a protocol for growing up cells and
inducing protein expression.

Protocol:
1. Single strains were plated on SD-URA agar and incubated at 30 C.
2. One colony (plate from step1) was inoculated in to 5 ml SD-URA liquid
  medium and incubated overnight at 30 C with shaking (200 rpm).
3. 1ml of the 5-ml culture from step 2, was inoculated into 25 ml -URA 2% 
  raffinose and incubated overnight at 30 C with shaking (200 rpm).
4. 200 ml of -URA 2% raffinose was inoculated with the culture from step 3 
  to a starting OD600nm ~ 0.1 and incubated at 30 C with shaking (200 rpm), 
  until an OD600nm ~ 1.2 - induction OD.


STRAIN  STARTING OD600           INDUCTION OD600

YGL202W 0.092 at time 0 hours    1.166 at time 21 hours
YJL060W 0.089 at time 0 hours    1.130 at time 18 hours
YER152C 0.101 at time 0 hours    1.142 at time 18 hours
YDL168W 0.107 at time 0 hours    1.136 at time 16.5 hours

5. 2 x 2ml samples were taken (when the cultures had reached induction
  OD) as a pre-induction blank before adding 3xYP 6% galactose.

6. Protein expression was induced by adding 100 ml 3xYP 6% galactose to
  the remainder of the culture from step 4, and the cultures incubated
  at 30 ºC with shaking (200 rpm).

7. Samples (2 x 2ml) were taken 2, 4 and 6 h from the start of
  induction process.

8. After 6 h, the cultures were harvested. The remaining cell
  suspension in the flask was divided in to 50 ml tubes and
  centrifuged at 4,000 rpm for 10 min at 4 C. The supernatant in each
  tube was replaced with 2 ml water, the yeast pellets were
  re-suspended and combined. Samples (1 ml) of this concentrated cell
  suspension were placed in 2-ml tubes and centrifuged at 5000 rpm for
  10 min at 4 C. The supernatant was removed and the cell pellets were
  stored frozen at -80 C.