Figure 3
Purification of PA2949 from P. aeruginosa. (a) P. aeruginosa PA01 cells carrying the pa2949 expression vector (pBBR-pa2949) or pBBR1mcs-3 (EV, empty vector) were harvested at an OD580 nm of 1 and disrupted. Membranes (M) were separated by ultracentrifugation. PA2949 was purified with an Ni–NTA column and subsequently with a UNOsphere Q anion-exchange column (AEx) in the presence of n-octyl β-D-glucoside. The samples were analyzed by SDS–PAGE stained with Coomassie Brilliant Blue G-250. The molecular weights of protein standards (St) are indicated on the right in kDa. |