Characterisation of a cDNA encoding gamma-glutamylcysteine synthetase in Medicago truncatula.
A gamma-ECS cDNA from Medicago truncatula was isolated using an Arabidopsis thaliana cDNA as probe. The analysis of the amino acid sequence deduced from this cDNA revealed 80% identity with the gamma-ECS from A. thaliana and Brassica juncea and suggested a plastidial localisation for the enzyme. Gamma-ECS activity and high level of GSH were detected in the gamma-ECS-deficient E. coli strain expressing a fusion protein containing the M. truncatula gamma-ECS protein. Southern blot analysis suggests that gamma-ECS is encoded by a small multigenic family in M. truncatula and shows that homologous genes are present in two other leguminous plants, Medicago sativa and Pisum sativum. Gamma-ECS gene expression was analysed by Northern blot in seedlings, plantlets and mature plants.
Frendo
P
P
Laboratoire de Biologie Végétale et Microbiologie, Centre National de la Recherche Scientifique, Université de Nice Sophia-Antipolis, France. frendo@unice.fr
Mathieu
C
C
Van de Sype
G
G
Hérouart
D
D
Puppo
A
A
D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification; D018744:DNA, Plant / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005721:Glutamate-Cysteine Ligase / Q000235:genetics*; D000455:Medicago sativa / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D018532:Pisum sativum / Q000201:enzymology / Q000235:genetics
D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D018744:DNA, Plant; D005721:Glutamate-Cysteine Ligase