A synthetic xylanase as a novel reporter in plants.
Transient gene expression assays are often used to screen promoters before stable transformation. Current transient quantification methods have several problems, including a lack of reporter gene stability and expense. Here we report a synthetic, codon-optimised xylanase gene ( sXynA) as a reporter gene for quantitative transient analyses in plants. Azurine-crosslinked xylan (AZCL-xylan) was used as a substrate for assaying xylanase activity. The enzymatic nature of the protein allows for sensitive assays at the low levels of transgene protein found in transiently transformed tissue extracts. The xylanase (XYN) protein is stable, activity slopes are linear over long time periods and assays are cost-effective. Coupled with the GUS Plus reporter gene, the XYN reporter allows sensitive and accurate quantification of gene control sequences in transient expression systems.
Vickers
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CSIRO Plant Industry, 306 Carmody Rd, St Lucia, 4067 Brisbane, QLD, Australia. claudia.vickers@csiro.au
Xue
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Gresshoff
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D000595:Amino Acid Sequence; D001483:Base Sequence; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D014990:Xylans / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D014990:Xylans; D043364:Endo-1,4-beta Xylanases