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GrainGenes Reference Report: JCS-33-143

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Reference	JCS-33-143
Title	Cloning and expression of a LMW-i glutenin gene
Journal	Journal of Cereal Science
Year	2001
Volume	33
Pages	143-154
Author	Cloutier S Rampitsch C Penner GA Lukow OM
Abstract	Summary: Endosperm-specific low-molecular-weight (LMW) glutenins are an important component of the polymeric gluten and, as such, play a key role in end-use functionality. Reports of N-terminal amino acid sequences of LMW glutenin fractions revealed that they have either a methionine or a serine residue at the first position of the mature peptide. These subunits were therefore called LMW-m and LMW-s type glutenins. A gene that is predicted to encode a LMW glutenin subunit having an isoleucine amino acid residue at position one of the mature protein was amplified and cloned from extra strong bread wheat cultivar Glenlea (pGH3.1). The predicted N-terminal sequence of this gene is truncated as compared to the m-type and s-type. The gene still codes for the expected number of eight cysteine residues which are all located in the C-terminal region. We propose to call it LMW-i based on the same nomenclature. Analysis of 277 doubled haploid lines derived from a single cross showed perfect co-segregation of the cloned PCR fragment with a rare LMW glutenin called LMW-50. The gene was subcloned in an expression vector and the protein was expressed in E. coli. Western blot analysis using a prolamin-specific monoclonal antibody confirmed the co-migration of the cloned protein with LMW-50 from Glenlea
Keyword	[ Hide all but 1 of 44 ] amino acid antibody bread bread wheat breadmaking quality cloning co-segregation analysis cross cysteine cysteine residues cystine peptides disulfide bonds doubled haploid doubled haploid lines durum wheat expression in e.coli food functionality gene gluten glutenin glutenin subunit haploid haploid lines lmw lmw glutenin genes lmw i methionine molecular-weight glutenin monoclonal antibodies n terminal sequence nomenclature pcr peptide primitive wheats residue serine subunit subunit proteins triticum durum vector western blot analysis wheat cultivar wheat storage proteins

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