Journal:

Article Title: The Arabidopsis thaliana proton transporters, AtNhx1 and Avp1, can function in cation detoxification in yeast

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Figure Lengend Snippet: Expression of Arabidopsis vacuolar pyrophosphatase AVP1 in ena1 mutants. (A) Vector pYES2 (Invitrogen) was introduced into wild-type, ena1, ena1 nhx1, and ena1 gef1 mutants. Plasmid pYes2-AVP1-D (13) was introduced into ena1, ena1 nhx1, and ena1 gef1 mutants. Five-fold serial dilutions (starting at 105 cells) of each strain were plated on YPGAL (1% yeast extract/2% peptone/2% galactose) with or without 0.5 M NaCl and incubated at 30°C for 2 days. (B and C) Intracellular concentrations of Na+ and K+. Exponentially growing cells (wild-type and ena1 transformed with pYES2 vector and ena1,ena1 nhx1, and ena1 gef1 mutants carrying pYes2-AVP1-D) were exposed to 0.7 M NaCl for 6 h. Total cell extracts were prepared (see Materials and Methods), and Na+ and K+ concentrations were determined. Values are the mean of two determinations, and bars represent the standard deviations. There is a consistent reduction in total cell Na+ in the ena1 AVP-D strain. The reason for this reduction is unknown.

Article Snippet: Cells were grown in YPD (1% yeast/2% peptone/2% dextrose; Difco), YPGAL (1% yeast/2% peptone/2% galactose; Difco), SD (Difco; Synthetic medium with 2% Dextrose), or APG (APG is a synthetic minimal medium containing 10 mM arginine, 8 mM phosphoric acid, 2% glucose, 2 mM MgSO 4 , 1 mM KCl, 0.2 mM CaCl 2 , and trace minerals and vitamins) ( 12 ).

Techniques: Expressing, Plasmid Preparation, Incubation, Transformation Assay