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Synthesis of a modified gene encoding human ornithine transcarbamylase for expression in mammalian mitochondrial and universal translation systems: a novel approach towards correction of a genetic defect
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posted on 2023-06-08, 14:44 authored by Vanessa C Wheeler, Chrisostomos ProdromouChrisostomos Prodromou, Laurence PearlLaurence Pearl, Robert Williamson, Charles CoutelleThe mitochondrial (MT) genome is a potential means of gene delivery to human cells for therapeutic expression. As a first step towards this, we have synthesized a gene coding for mature human ornithine transcarbamylase (OTC) by recursive PCR using 18 oligodeoxyribonucleotides, each 70-80 nucleotides in length, using codons which should allow translation in accordance with both mammalian mt and universal codon usage. Flanking mt DNA sequences were incorporated which are designed to facilitate site-specific cloning into the mt genome. Expression of this human gene in Escherichia coli leads to an immunoreactive OTC product of the correct size and N-terminal amino-acid sequence, but which forms inclusion bodies and lacks enzymatic activity.
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Publication status
- Published
Journal
GeneISSN
0378-1119Publisher
ElsevierExternal DOI
Issue
2Volume
169Page range
251-255Department affiliated with
- Biochemistry Publications
Full text available
- No
Peer reviewed?
- Yes
Legacy Posted Date
2015-02-24Usage metrics
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