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GrainGenes Reference Report: PNA-97-1914

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Reference	PNA-97-1914
Title	The production of recombinant proteins in transgenic barley grains
Journal	Proceedings of the National Academy of Sciences, USA
Year	2000
Volume	97
Pages	1914-1919
Author	Horvath H [ Show all 7 ]
Abstract	Summary: The grain of the self-pollinating diploid barley species offers two modes of producing recombinant enzymes or other proteins. One uses the promoters of genes with aleurone-specific expression during germination and the signal peptide code for export of the protein into the endosperm. The other uses promoters of the structural genes for storage proteins deposited in the developing endosperm. Production of a protein-engineered thermotolerant (1, 3-1, 4)-beta-glucanase with the D hordein gene (Hor3-1) promoter during endosperm development was analyzed in transgenic plants with four different constructs. High expression of the enzyme and its activity in the endosperm of the mature grain required codon optimization to a C+G content of 63% and synthesis as a precursor with a signal peptide for transport through the endoplasmic reticulum and targeting into the storage vacuoles. Synthesis of the recombinant enzyme in the aleurone of germinating transgenic grain with an alpha-amylase promoter and the code for the export signal peptide yielded approximately equal to 1 microgram(.)mg-1 soluble protein, whereas 54 microgram(.)mg-1 soluble protein was produced on average in the maturing grain of 10 transgenic lines with the vector containing the gene for the (1, 3-1, 4)-beta-glucanase under the control of the Hor3-1 promoter
Keyword	[ Hide all but 1 of 44 ] agrobacterium tumefaciens aleurone alpha amylase barley barley grain beta glucanase codon optimization development endoplasmic reticulum endosperm enzyme gene gene expression gene transfer genetic transformation germination grains heat stability hordein hordein gene hordeum vulgare its optimization organelles peptide plasmid vectors precursor promoter recombinant dna recombinant protein signal signal peptide storage storage protein storage vacuoles structural genes synthesis targeting transgenic transgenic barley transgenic plant transport vacuole vector

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