Molecular cloning and characterisation of two genes encoding asparagine synthetase in barley (Hordeum vulgare L.)

TY - JOUR

T1 - Molecular cloning and characterisation of two genes encoding asparagine synthetase in barley (Hordeum vulgare L.)

AU - Møller, Marianne G.

AU - Taylor, Christopher

AU - Rasmussen, Søren K.

AU - Holm, Preben B.

PY - 2003/7/28

Y1 - 2003/7/28

N2 - Two different cDNA clones encoding asparagine synthetase (AS: EC 6.3.5.4.) were cloned from barley (Hordeum vulgare L. cv. Alexis). The corresponding genes were designated HvAS1 (GenBank no AF307145) and HvAS2 (GenBank no AY193714). Chromosomal mapping using wheat-barley addition lines revealed that the HvAS1 gene is located on the long arm of barley chromosome 5, while the HvAS2 gene maps to the short arm of chromosome 3. Both genes are expressed in barley leaves according to RT-PCR analysis but only the HvAS1 gene expression can be detected in roots. Northern blots show no expression of HvAS1 in plants grown under a normal 16 h light/8 h dark cycle but after 10 h of continuous darkness, transcript appears and mRNA accumulates over a 48-h period of dark treatment. In roots, low-level expression of HvAS1 could be detected and the expression level appears to be unaffected by light. A polyclonal antibody was raised against the HvAS1 protein and used in Western blot analysis. The AS protein accumulated during a 48-h period of dark treatment, following the increase in HvAS1 transcript.

AB - Two different cDNA clones encoding asparagine synthetase (AS: EC 6.3.5.4.) were cloned from barley (Hordeum vulgare L. cv. Alexis). The corresponding genes were designated HvAS1 (GenBank no AF307145) and HvAS2 (GenBank no AY193714). Chromosomal mapping using wheat-barley addition lines revealed that the HvAS1 gene is located on the long arm of barley chromosome 5, while the HvAS2 gene maps to the short arm of chromosome 3. Both genes are expressed in barley leaves according to RT-PCR analysis but only the HvAS1 gene expression can be detected in roots. Northern blots show no expression of HvAS1 in plants grown under a normal 16 h light/8 h dark cycle but after 10 h of continuous darkness, transcript appears and mRNA accumulates over a 48-h period of dark treatment. In roots, low-level expression of HvAS1 could be detected and the expression level appears to be unaffected by light. A polyclonal antibody was raised against the HvAS1 protein and used in Western blot analysis. The AS protein accumulated during a 48-h period of dark treatment, following the increase in HvAS1 transcript.

KW - Asparagine synthetase

KW - C/N ratio

KW - Chromosomal localisation

KW - Gene expression

KW - Nitrogen metabolism

UR - http://www.scopus.com/inward/record.url?scp=0142043940&partnerID=8YFLogxK

U2 - 10.1016/S0167-4781(03)00137-4

DO - 10.1016/S0167-4781(03)00137-4

M3 - Journal article

C2 - 12890559

AN - SCOPUS:0142043940

SN - 0167-4781

VL - 1628

SP - 123

EP - 132

JO - Biochimica et Biophysica Acta, Gene Structure and Expression

JF - Biochimica et Biophysica Acta, Gene Structure and Expression

IS - 2

ER -