Expression of murine adenosine deaminase (ADA) in transgenic maize.
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.
Petolino
J F
JF
Dow AgroSciences, Indianapolis, IN 46077, USA. JFPetolino@DowAgro.com
Young
S
S
Hopkins
N
N
Sukhapinda
K
K
Woosley
A
A
Hayes
C
C
Pelcher
L
L
D000241:Adenosine / Q000494:pharmacology; D000243:Adenosine Deaminase / Q000235:genetics* / Q000378:metabolism; D000613:Aminobutyrates / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D003839:Deoxyadenosines / Q000494:pharmacology; D006540:Herbicides / Q000494:pharmacology; D051379:Mice; D030821:Plants, Genetically Modified; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D012639:Seeds / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
D000613:Aminobutyrates; D003839:Deoxyadenosines; D006540:Herbicides; C003121:phosphinothricin; D000243:Adenosine Deaminase; D000241:Adenosine; C058118:2'-deoxyadenosine