摘要
本研究利用大肠杆菌的 GUS 基因(β-葡糖苷酸酶基因,为报告基因,以改良的 PEG 转化方法),对大豆(Glycine max L.)未成熟子叶原生质体进行直接转化。在原生质体培养基 KP8中培养1—6天和14—30天后,利用 GUS 基因产物与底物 MUG 反应的荧光分析和与底物X-Gluc 反应的组织化学定位实验,分别检测到 GUS 基因在大豆未成熟子叶原生质体中的瞬间和稳定表达。同时,对影响 PEG 转化的渚多因素,如 PEG 的毒性、转化后的原生质体稳定性等进行了讨论。
The E.coli beta-glucuronidase (GUS) gene was used in gene expression experiments with the protoplasts isolated from immature cotyledons of soybean (Glycine max L.).Transient expre- ssion of GUS gene could be detected in 1—6 days after DNA incorporation into soybean proto- plasts by using a fluorogenic substrate MUG.Stable transformation was observed by histoche- mical localization with the use of a substrate X-Gluc.Some factors such as PEG toxicity and protoplast stability affecting PEG-mediated transformation were discussed.
基金
"七.五"生物工程攻关项目资助
关键词
大豆
原生质体
瞬间表达
GUS基因
Cultivated soybean(Glycine max L.)
Protoplasts
Transient expression
Transformation
GUS gene
