Skip to main content
. 2006 Apr;140(4):1279–1296. doi: 10.1104/pp.105.075150

Figure 4.

Figure 4.

Bacterial expression and western-blot analysis of various PcINO1 mutants, Os∷PcINO1, BjINO1, and Bj∷PcINO1 gene products, their in vitro MIPS activity, and salt tolerance vis à vis salt sensitivity. A and C, Ten-percent SDS-PAGE. B and D, Corresponding western-blot analysis of PcINO1 mutants, Os∷PcINO1, BjINO1, and Bj∷PcINO1, respectively. A and B, Lane 1, Mr marker; lanes 2 to 5, pellet fractions of overexpressed PcINO1, ΔPcINO1.1, ΔPcINO1.2, and Os∷PcINO1, respectively; lanes 6 to 9, corresponding supernatants, respectively. C and D, Lane 1, Mr marker; lane 2, OsINO1 (+ control); lanes 3 and 4, induced BjINO1 supernatant and pellet fractions, respectively; lanes 5 and 6, induced Bj∷PcINO1 supernatant and pellet fractions, respectively. E, In vitro MIPS activity of purified PcINO1 mutants, Os∷PcINO1, BjINO1, and Bj∷PcINO1 along with the positive controls PcINO1 and OsINO1. Specific activity is described in terms of μmol inositol 1-P h−1 mg protein−1. In each case 20 μg purified protein was taken. Error bars indicate the deviation of triplicate experiments. F, Effect of increasing concentration of NaCl on the MIPS activity of ΔPcINO1.2 (white circle), Os∷PcINO1 (black triangle), BjINO1 (white square), Bj∷PcINO1 (white triangle), along with OsINO1 (−control, black square) and PcINO1 (+control, black circle). Approximately 20 μg of each purified enzyme was assayed in presence of indicated concentration of NaCl. Error bars indicate the deviation from triplicate experiments.