TABLE 2.
Protein yielda
| Fraction | Yield (pmol/g [wet wt] of cells)
|
||||
|---|---|---|---|---|---|
| JY741/f-Rpb3
|
JY741/f-Fcp1
|
||||
| Rpb1 | Fcp1 | Rpb1 | f-Fcp1 | ||
| Supernatant of low-speed centrifugation | 37 | 10 | 33 | 7.6 | |
| First extract | 16 | 9.9 | 16 | 8.0 | |
| M2-agarose-bound fraction | 5.0 | 2.7 | 1.7 | 2.4 | |
| Wash fraction | 5.6 | 1.0 | 3.1 | 0.65 | |
| Nuclease-treated extract | 0.61 | 0.15 | 0.57 | 0.11 | |
| M2-agarose-bound fraction | 0.12 | 0.04 | 0.04 | 0.03 | |
| Final precipitateb | >22 | >7.9 | >11 | >4.6 | |
a
One gram of cells corresponds to 6 × 109 cells. The purification was repeated using different amounts of cells, and a typical result is presented.
b
Rpb1 detected in the final precipitate was mostly in the IIo form (data not shown).