cDNA cloning and characterization of the novel genes related to aldehyde dehydrogenase from wild Chinese grape (Vitis pseudoreticulata W. T. Wang).

mRNA differential display was employed to study the gene differential expression of wild Chinese grape (Vitis pseudoreticulata W. T. Wang) infected by Uncinula necator in different periods, a cDNA fragment of T11AC/B0319-456 coded by aldehyde dehydrogenase (ALDH) gene has been obtained. 5' RACE and 3' RACE have been used to clone the whole cDNA sequences of ALDH which consists of three cDNA sequences, whose sizes are 1887, 1956 and 1961 bp, and they encoded a polypeptide size of 537, 524 and 477 designated as VpALDH2a, VpALDH2b and VpALDH1a, respectively. The deduced amino acid sequence shared highly identity with other plants and Human ALDH. Both VpALDH2a and VpALDH2b protein contain putative mitochondrial targeting sequence except VpALDH1a, it indicates that VpALDH2a and VpALDH2b are mitochondrial enzymes, and VpALDH1a is cytosolic enzyme. The VpALDH2a was subcloned into the expression vector pGEX-4T-1, transformed into E.coli BL 21-coden plus induced by IPTG, and about Mr. 85 kD of GST-ALDH fusion protein displayed in SDS-PAGE gel.