Construction and expression of a synthetic wheat storage protein gene

A synthetic wheat high-molecular-weight (HMW) glutenin storage protein gene analog was constructed for expression in E. coli. This first synthetic HMW-glutenin gene and future modifications are intended to allow systematic dissection of the molecular basis of HMW-glutenin role in the visco-elastic p...

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Bibliographic Details
Published in:Gene 1996-09, Vol.174 (1), p.51-58
Main Authors: Anderson, Olin D., Kuhl, Joseph C., Tam, Angie
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacterial expression
Base Sequence
codon usage
codons
dna modification
Escherichia coli
Escherichia coli - genetics
gene expression
gene transfer
Genes, Plant
Genes, Synthetic
genetic code
genetic transformation
glutenins
Glutens - analogs & derivatives
Glutens - genetics
high molecular weight glutenins
High-molecular-weight glutenin
Molecular Sequence Data
nucleotide sequences
Oligonucleotides
pET3a vector
Polymerase-chain-reaction
Repetitive domain
repetitive sequence domains
repetitive sequences
Repetitive Sequences, Nucleic Acid
restriction endonucleases
restriction site sequences
synthesis
Synthetic gene
synthetic genes
Triticum - genetics
Triticum aestivum
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Summary:A synthetic wheat high-molecular-weight (HMW) glutenin storage protein gene analog was constructed for expression in E. coli. This first synthetic HMW-glutenin gene and future modifications are intended to allow systematic dissection of the molecular basis of HMW-glutenin role in the visco-elastic properties critical for wheat product processing and utilization. The design of the gene included four features: different construction strategies for the separate assembly of major polypeptide domains, the inclusion of convenient restriction sites for modifications, use of a codon selection similar to E. coli highly expressed genes, and the ability to produce repetitive sequence domains of exact numbers of defined repeats. The complete synthetic HMW-glutenin construct was 1908 bp, and contained 32 identical copies of one of the HMW-glutenin repetitive domain motifs. The gene expressed the novel HMW-glutenin protein to relatively high levels in bacterial cultures and the protein exhibited the known anomalous behavior of HMW-glutenins in SDS-PAGE.
ISSN:0378-1119
1879-0038
DOI:10.1016/0378-1119(96)00315-0