Expression of murine adenosine deaminase (ADA) in transgenic maize

Transgenic Res. 2000 Feb;9(1):1-9. doi: 10.1023/a:1008972101370.

Abstract

A murine adenosine deaminase (ADA) gene, driven by the maize ubi-1 promoter and intron region, was transformed into embryogenic maize callus, along with a bar and gusA gene-containing plasmid, using microparticle bombardment. Selection in the presence of either the herbicide Basta or the adenosine analogue 2'-deoxyadenosine resulted in transgenic cultures that expressed GUS and accumulated a 41-kD protein that immunoprecipated with an ADA-specific polyclonal antibody. ADA enzyme activity was observed in extracts from transgenic callus as well as regenerated plants and progeny. Culltures expressing ADA grew in the presence of 200 mg/l 2'-deoxyadenosine, a concentration which completely inhibited the growth of non-transgenic cultures. ADA activity appeared to segregate in progency of regenerated plants as a single, dominant Mendelian trait. These results suggest that ADA, in combination with adenosine analogue selection, represents a potentially viable selectable marker system for transgenic maize production.

MeSH terms

  • Adenosine / pharmacology
  • Adenosine Deaminase / genetics*
  • Adenosine Deaminase / metabolism
  • Aminobutyrates / pharmacology
  • Animals
  • Blotting, Western
  • Deoxyadenosines / pharmacology
  • Herbicides / pharmacology
  • Mice
  • Plants, Genetically Modified
  • Precipitin Tests
  • Promoter Regions, Genetic
  • Seeds / genetics
  • Seeds / metabolism
  • Zea mays / enzymology
  • Zea mays / genetics*
  • Zea mays / metabolism

Substances

  • Aminobutyrates
  • Deoxyadenosines
  • Herbicides
  • phosphinothricin
  • Adenosine Deaminase
  • Adenosine
  • 2'-deoxyadenosine