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• W259134615 abstract "The young internodes of sugarcane are ideal targets for altering metabolism, through genetic manipulation, to potentially control known fungal diseases such as Smut or to increase sucrose yields in these regions that are currently being discarded. At present, no regulatory sequences that specifically drive transgene expression in young developing sugarcane tissues are available. The objective of this study was therefore to isolate and evaluate such a sequence. The promoter targeted for isolation in this study regulates the expression of UDP-glucose dehydrogenase (EC 1.1.1.22), an enzyme which catalyses the oxidation of UDP-glucose to UDP-glucuronic acid, a precursor for structural polysaccharides which are incorporated into the developing cell wall. A strong correlation between the expression of UDP-glucose dehydrogenase and a demand for structural polysaccharides in developing tissues could therefore be expected. The first part of this study addressed the general practicality of promoter isolation from sugarcane, a complex polyploid. A gene encoding UDP-glucose dehydrogenase was isolated from a sugarcane genomic library. The gene contains an open reading frame (ORF) of 1443 bp, encoding 480 amino acids and one large intron (973 bp), located in the 5’-UTR. The derived amino acid sequence showed 88 – 98% identity with UDP-glucose dehydrogenase from other plant species, and contained highly conserved amino acid motifs required for cofactor binding and catalytic activity. Southern blot analysis indicates a low copy number for UDP-glucose dehydrogenase in sugarcane. The possible expression of multiple gene copies or alleles of this gene was investigated through comparison of sequences amplified from cDNA prepared from different tissues. Although five Single Nucleotide Polymorphisms (SNP) and one small-scale insertion/deletion (INDEL) were identified in the aligned sequences, hundred percent identity of the derived amino acid sequences suggested the expression of different alleles of the same gene rather than expression of multiple copies. The finding that multiple alleles are expressed to provide the required level of a specific enzyme, rather than the increased expression of one dominant allele, is encouraging for sugarcane gene and promoter isolation. In the second part of the study the suitability of UDP-glucose dehydrogenase as a target for the isolation of a developmentally regulated promoter was investigated. The contribution of UDP-" @default.
• W259134615 created "2016-06-24" @default.
• W259134615 creator A5030382391 @default.
• W259134615 date "2006-12-01" @default.
• W259134615 modified "2023-09-27" @default.
• W259134615 title "Isolation and evaluation of the sugarcane UDP-glucose dehydrogenase gene and promoter" @default.
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