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. 2006 Sep;128(3):283–292. doi: 10.1085/jgp.200609572

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Copyright © 2006, The Rockefeller University Press

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Figure 2. — Expression, purification, and functional reconstitution of KvLm in proteoliposomes. (A) Immunoblots of E. coli cell lysates: lane 1, M_r standards; lanes 2 and 3, uninduced (UI) and IPTG-induced (I) cultures transformed with pQE-70 containing the gene encoding KvLm; lanes 4 and 5, UI and I control cultures transformed with empty vector. A prominent band on lane 3 is at an anticipated M_r ∼29 kD for KvLm. Lane 6, immunoblot of purified KvLm; the 29 kD protein retains propensity to oligomerize in SDS. (B) Time course of ⁸⁶Rb⁺ uptake by KvLm reconstituted in proteoliposomes at 10 μg protein/mg phospholipid. (C) Ionic selectivity of KvLm-evoked ⁸⁶Rb⁺ uptake. Bars represent uptake at 30 min into proteoliposomes prepared in 0.45 M Cl⁻ salts of the indicated cations.