Heterologous expression and purification of native and mutated low molecular mass glutenin subunits from durum wheat.
Wheat technological properties are correlated with the size of glutenin polymers, consisting of high and low molecular mass glutenin subunits, linked together by disulphide bonds. In order to unravel glutenin polymer structure, we considered three LMW-GS genes, which differ in the number of cysteine residues and in the repetitive domain length. The three LMW-GS genes have been expressed in Escherichia coli, and purified with a yield of 40-100 mg/l of culture volume, depending on protein type. Single polypeptides are being used in re-oxidation and micro-mixographic experiments, in order to detect the influence of the differential structural characteristics on glutenin polymer formation.
Patacchini
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Dipartimento di Agrobiologia e Agrochimica, Università degli Studi della Tuscia, Via S. Camillo de Lellis, 01100, Viterbo, Italy. cpatacchini@unitus.it
Masci
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D'Ovidio
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Lafiandra
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D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005983:Glutens / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D008970:Molecular Weight; D009154:Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014908:Triticum / Q000737:chemistry*
D017931:DNA Primers; D011994:Recombinant Proteins; D005983:Glutens; C037302:glutenin