Molecular characterization of the glnA gene encoding glutamine synthetase from the edible mushroom Agaricus bisporus

Abstract

The gene encoding glutamine synthetase (glnA) was isolated from an Agaricus bisporus H39 recombinant k phage library. The deduced A. bisporus glutamine synthetase amino acid sequence contains 354 residues.

Expression of the urease gene of Agaricus bisporus: a tool for studying fruit body formation and post-harvest development

“…The decrease in urea content of the growing sporophore was thus not only the result of the dilution effect as the fruit body develops and grows to a larger volume. The urease activity demonstrated indicates that ammonium is produced and can be reassimilated by the primary ammonium assimilation pathways addressed by Baars et al (1996) and Kersten et al (1997Kersten et al ( , 1998Kersten et al ( , 1999. For mature fruit bodies of A. bisporus, however, no reassimilation of urea-ammonia was determined as was shown for puffballs of Lycoperdon pyriforme (Reinbothe et al 1967).…”

“…The PCR product of forward primer 5′-GGC GAC GAA GTC AAA TTT GG-3′ (ure1f) and reverse primer 5′-TCG TTG AGA GTG TCG GTATG-3′ (ure2r) was labelled with [ 32 P] CTP and used as a probe (Wagemaker et al 2005). Northern blots were also probed with an A. bisporus 28S ribosomal DNA fragment as loading control and with probes specific for the genes gdhA, gdhB and glnA encoding enzymes of the primary nitrogen metabolism (Kersten et al 1997(Kersten et al , 1999.…”

“…The slight increase at 15-18 h is not significant when the signals are corrected for the 28S rRNA loading control. For comparison, the blot was also hybridized with probes specific for the NADP-dependent GDHs (gdhA, gdhB) and the glutamine synthetase (glnA) of A. bisporus (Kersten et al 1997(Kersten et al , 1999. These enzymes of the primary nitrogen metabolism showed the same expression pattern (Fig.…”

Classification of Sequences Expressed during the Primordial and Basidiome Stages of the Cultivated Mushroom Agaricus bisporus

“…Previous genomic studies of A. bisporus have focused on the cloning and characterization of individual genes of significant metabolic importance. Genes encoding glyceraldehyde-3-phosphate dehydrogenase (Harmsen et al, 1992), NADP ϩ -dependent glutamate dehydrogenase , the ␦ subunit of the mitochondrial ATP synthase (De Groot et al, 1997), hydrophobins A, B, andC (De Groot et al, 1996), glutamine synthetase (Kersten et al, 1997), laccase (Smith et al, 1998), and endo-1,4-␤-xylanase (De Groot et al, 1998a) successfully were characterized. Only the hydrophobin-and the ATP-synthase ␦-subunit-encoding genes, as well as several uncharacterized transcripts, appear to be differentially expressed during A. bisporus development, as suggested by Northern blot analyses (De Groot et al, 1997).…”

NAD+-dependent glutamate dehydrogenase of the edible mushroom Agaricus bisporus: biochemical and molecular characterization

“…For DNA isolation, the strains were cultured according to Baars et al (1994) on DT80 medium supplemented with glucose as the carbon source. For enzyme measurements and Northern analyses, mycelium of strain Horst U1 was grown as described by Kersten et al (1997) on the media described in the Results section. These cultures were either analyzed directly or used for shift experiments as described before (Kersten et al 1997).…”