membrane potentials (ΔΨ) of –100 – –150 mV inside. These values are physiologically significant, as several membranes in plants maintain ΔΨ in this range. However, the author had difficulty reproducing these results due to an apparent "death" of oocytes containing the AtMRS2-11 protein, and therefore these results should be viewed with caution. The AtMRS2-10 transporter has been analysed using radioactive tracer uptake analysis. 63Ni2+ was used as the substitute ion and Mg2+ was shown to inhibit the uptake of 63Ni2+ with a Ki of 20 μM. Uptake was also inhibited by Co(III)Hex and by other divalent cations. Only Co2+ and Cu2+ inhibited transport with Ki values less than 1 mM. The AtMRS2-10 protein was fused to GFP, and was shown to be localised to the plasma membrane. A similar experiment was attempted in the Schock et al. (2000) paper, but the observed localisation was not significantly different from that seen with unfused GFP. The most likely reason for the lack of a definitive localisation of AtMRS2-1 in the Schock et al. paper is that the authors removed the TM domains from the protein, thereby precluding its insertion into a membrane. The exact physiological significance of the AtMRS2-1 and AtMRS2-10 proteins in plants has yet to be clarified. The AtMRS2-11 gene has been overexpressed (from the CaMV 35S promoter) in A. thaliana. The transgenic line has been shown to accumulate high levels of the AtMRS2-11 transcript. A strong Mg2+ deficiency phenotype (necrotic spots on the leaves, see Chapter 1.5 below) was recorded during the screening process (in both the T1 and T2 generations) for a homozygote line, but this phenotype was lost in the T3 generation and could not be reproduced when the earlier generations were screened a second time. The author suggested that environmental effects were the most likely cause of the inconsistent phenotype. AtMHX The first magnesium transporter isolated in any multicellular organism, AtMHX shows no similarity to any previously isolated Mg2+ transport protein. The gene was initially identified in the A. thaliana genomic DNA sequence database, by its similarity to the SLC8 family of Na+/Ca2+ exchanger genes in humans. The cDNA sequence of 1990 bp is predicted to produce a 539-amino acid protein. AtMHX is quite closely related to the SLC8 family at the amino acid level and shares a topology with eleven predicted TM domains (Figure A10.5). There is one major difference in the sequence, in that