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Biotech Histochem. 2015;90(6):453-60. doi: 10.3109/10520295.2015.1020875. Epub 2015 Apr 29.

Gene transcript accumulation and in situ mRNA hybridization of two putative glutamate dehydrogenase genes in etiolated Glycine max seedlings.

Author information

1
Laboratory of General and Agricultural Microbiology, Agricultural University of Athens , Athens , Greece.

Abstract

Glutamate dehydrogenase (EC 1.4.1.2) is a multimeric enzyme that catalyzes the reversible amination of α-ketoglutarate to form glutamate. We characterized cDNA clones of two Glycine max sequences, GmGDH1 and GmGDH2, that code for putative α- and β-subunits, respectively, of the NADH dependent enzyme. Temporal and spatial gene transcript accumulation studies using semiquantitative RT-PCR and in situ hybridization have shown an overlapping gene transcript accumulation pattern with differences in relative gene transcript accumulation in the organs examined. Detection of NADH-dependent glutamate dehydrogenase activity in situ using a histochemical method showed concordance with the spatial gene transcript accumulation patterns. Our findings suggest that although the two gene transcripts are co-localized in roots of etiolated soybean seedlings, the ratio of the two subunits of the active holoenzyme may vary among tissues.

KEYWORDS:

Glycine max; NADH glutamate dehydrogenase; RT-PCR; in situ hybridization; root

PMID:
25922975
DOI:
10.3109/10520295.2015.1020875
[Indexed for MEDLINE]

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