Petolino J F, Young S, Hopkins N, Sukhapinda K, Woosley A, Hayes C, Pelcher L
Dow AgroSciences, Indianapolis, IN 46077, USA.
Transgenic Res. 2000 Feb;9(1):1-9. doi: 10.1023/a:1008972101370.
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.
一个由玉米泛素-1启动子和内含子区域驱动的小鼠腺苷脱氨酶(ADA)基因,与一个含有bar和gusA基因的质粒一起,通过微粒轰击法转化到胚性玉米愈伤组织中。在除草剂Basta或腺苷类似物2'-脱氧腺苷存在的情况下进行筛选,得到了表达GUS并积累了一种41-kD蛋白质的转基因培养物,该蛋白质能与ADA特异性多克隆抗体发生免疫沉淀反应。在转基因愈伤组织以及再生植株和后代的提取物中都观察到了ADA酶活性。表达ADA的培养物在200 mg/l 2'-脱氧腺苷存在的情况下生长,而该浓度完全抑制了非转基因培养物的生长。ADA活性在再生植株的后代中似乎作为一个单一的显性孟德尔性状分离。这些结果表明,ADA与腺苷类似物筛选相结合,代表了一种用于转基因玉米生产的潜在可行的选择标记系统。