Molecular phylogeny and evolution of alcohol dehydrogenase (Adh) genes in legumes

BACKGROUND: Nuclear genes determine the vast range of phenotypes that are responsible for the adaptive abilities of organisms in nature. Nevertheless, the evolutionary processes that generate the structures and functions of nuclear genes are only now be coming understood. The aim of our study is to isolate the alcohol dehydrogenase (Adh) genes in two distantly related legumes, and use these sequences to examine the molecular evolutionary history of this nuclear gene. RESULTS: We isolated the expressed Adh genes from two species of legumes, Sophora flavescens Ait. and Wisteria floribunda DC., by a RT-PCR based approach and found a new Adh locus in addition to homologues of the Adh genes found previously in legumes. To examine the evolution of these genes, we compared the species and gene trees and found gene duplication of the Adh loci in the legumes occurred as an ancient event. CONCLUSION: This is the first report revealing that some legume species have at least two Adh gene loci belonging to separate clades. Phylogenetic analyses suggest that these genes resulted from relatively ancient duplication events

Immunohistochemical and Semiquantitative Immunoblot Analyses of Nm23-Hl and H2 Isoforms in Normal Human Tissues

The total amount of nm23 protein, the relative ratios of H1 and H2 isoforms (H2/H1) and the localization of these proteins in human normal tissues were studied by a semiquantitative immunoblot technique followed by densitometry and immunohistochemistry with monoclonal antibody against nm23 protein (Pan-242). All tissues contained both isoforms recognized as the 20.5kD H1 protein and 18.5kD H2 protein by immunoblotting. Nm23 protein was abundant in liver, kidney and adrenal gland tissue, and scarce in heart and muscle. H2 levels were always higher than H1, but the isoform ratios (H2/H1) were variable from tissue to tissue. Immunostaining revealed that nm23 protein was predominantly present in cytoplasm and the pattern of staining was homogeneous in parenchymal cells of the liver, pancreas and colonic mucosa and heterogeneous in gastric mucosa and kidney. These results demonstrated that the levels of nm23 protein and the H2/H1 ratios and distribution of isoforms were different in each tissue, and suggests that, when the alterations of nm23 gene expression in tumor tissues are examined, the levels and ratios in non-tumorous tissues surrounding the tumor nest should be considered

Immunohistochemical and Semiquantitative Immunoblot Analyses of Nm23-Hl and H2 Isoforms in Normal Human Tissues

The total amount of nm23 protein, the relative ratios of H1 and H2 isoforms (H2/H1) and the localization of these proteins in human normal tissues were studied by a semiquantitative immunoblot technique followed by densitometry and immunohistochemistry with monoclonal antibody against nm23 protein (Pan-242). All tissues contained both isoforms recognized as the 20.5kD H1 protein and 18.5kD H2 protein by immunoblotting. Nm23 protein was abundant in liver, kidney and adrenal gland tissue, and scarce in heart and muscle. H2 levels were always higher than H1, but the isoform ratios (H2/H1) were variable from tissue to tissue. Immunostaining revealed that nm23 protein was predominantly present in cytoplasm and the pattern of staining was homogeneous in parenchymal cells of the liver, pancreas and colonic mucosa and heterogeneous in gastric mucosa and kidney. These results demonstrated that the levels of nm23 protein and the H2/H1 ratios and distribution of isoforms were different in each tissue, and suggests that, when the alterations of nm23 gene expression in tumor tissues are examined, the levels and ratios in non-tumorous tissues surrounding the tumor nest should be considered