The Construction of Halo-Tolerant Higher Plants which were Transformed by Marine Alga Na^+-ATPase gene.

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The ability which can export intracellular sodium ions can be transformed usual higher plants to halotolerant. The Na^+ K^+-ATPase of animal cells or Na^+-ATPase of raphidophycean H. akashiwo was able to transport Na ions. The Na^+-ATPase of H. akasiwo was consequently proper enzyme for the transformation. A DNA between conserved parts of eucaryotic ATPase was amplified from H. akasiwo cDNA library with PCR method. The Na^+-ATPase of H. akashiwo was cloned with the amplified DNA as a probe. Two kinds of ATPase molecules was cloned consequently. The amino acid sequences and the secondary structures analysis showed the homology to other species' ATPases(70% : protozoan L. donovani, 40-50% : higer plants and fungi, 20-30% : higher animals). This suggests that the ATPase amino acid sequences tend to conserve among systematical close to species rather than the functional characters (ion selectivity, etc). Rescreeing was needed to the reason why the two cDNA clones were short of complete coding regions. The most expressing cell period of the genes was determined with northern hybridization and 5' end probe was successed to get the enough length cDNA (3.4kb).