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Gene specificity is maintained in transient expression assays with protoplasts derived from different tissues of barley

Summary

In some cereal species that are still recalcitrant to stable transformation and regeneration, transient expression in isolated protoplasts is a useful tool for the study of gene expression and regulation. We have successfully applied these techniques to barley protoplasts derived from developing endosperm, aleurone, leaves and roots in order to characterize functionally cis-acting motives in two gene promoters, corresponding to trypsin inhibitor BTI-CMe and to sucrose synthase Ss1. Gene specificity is maintained in transient expression assays with protoplasts isolated from these different barley tissues and the pattern of expression parallels the mRNA levels observed for the corresponding genes in the same tissues.

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Abbreviations

CaMV35S:

35S-cauliflower mosaic virus promoter

GUS:

β-glucuronidase

MU:

4-methyl umbelliferone

NOS:

nopaline synthase gene

PEG:

polyethyleneglycol

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Díaz, I., Royo, J., de la Hoz, P.S. et al. Gene specificity is maintained in transient expression assays with protoplasts derived from different tissues of barley. Euphytica 85, 203–207 (1995). https://doi.org/10.1007/BF00023949

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Key words

  • barley
  • electroporation
  • PEG-mediated DNA uptake
  • promoter analysis
  • protoplasts
  • transient expression