A synthetic xylanase as a novel reporter in plants View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2003-09

AUTHORS

C. E. Vickers, G. P. Xue, P. M. Gresshoff

ABSTRACT

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. More... »

PAGES

135-140

References to SciGraph publications

  • 1987-12. Assaying chimeric genes in plants: The GUS gene fusion system in PLANT MOLECULAR BIOLOGY REPORTER
  • 1990-08. Characterization of the rice (Oryza sativa) actin gene family in PLANT MOLECULAR BIOLOGY
  • 1994-07. Quantitative transient gene expression: Comparison of the promoters for maize polyubiquitin1, rice actin1, maize-derivedEmu andCaMV 35S in cells of barley, maize and tobacco in TRANSGENIC RESEARCH
  • 1991-10. Sequence of three members and expression of a new major subfamily of glutelin genes from rice in PLANT MOLECULAR BIOLOGY
  • 1996-05. Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants in TRANSGENIC RESEARCH
  • 1994-11. Non-systemic expression of a stress-responsive maize polyubiquitin gene (Ubi-1) in transgenic rice plants in PLANT MOLECULAR BIOLOGY
  • 1993-11. Activity of a maize ubiquitin promoter in transgenic rice in PLANT MOLECULAR BIOLOGY
  • 1998-11. Identification of promoter elements in a low-temperature-responsive gene (blt4.9) from barley (Hordeum vulgare L.) in PLANT MOLECULAR BIOLOGY
  • 1992-02. Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation in PLANT MOLECULAR BIOLOGY
  • 2003-07. Selectable marker-free transgenic barley producing a high level of cellulase (1,4-β-glucanase) in developing grains in PLANT CELL REPORTS
  • 1992-07. Development of the particle inflow gun for DNA delivery to plant cells in PLANT CELL REPORTS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00299-003-0667-9

    DOI

    http://dx.doi.org/10.1007/s00299-003-0667-9

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1008499779

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/12845475


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    43 schema:description 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.
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