Laboratory of Gene Technology, Catholic University of Leuven, Willem de Croylaan 42, B-3001, Leuven, Belgium.
Plant Cell Rep. 1994 Feb;13(5):262-6. doi: 10.1007/BF00233316.
Electroporation conditions were established for transient expression of introduced DNA in banana (Musa spp., cv. 'Bluggoe') protoplasts isolated from regenerable embryogenic cell suspensions. The following parameters were found to be highly influential: electroporation buffer, polyethylene glycol treatment and its duration before electroporation, use of a heat shock, and chimaeric gene constructs. The maximum frequency of DNA introduction as detected by an in situ assay for transient expression of the uidA gene, amounted to 1.8% of total protoplasts. Since plants have recently been regenerated from banana protoplasts at a high frequency, the present results may contribute to the production of transgenic banana.
电穿孔条件为瞬时表达在香蕉(Musa spp.,cv. 'Bluggoe')原生质体中引入的 DNA 而建立,这些原生质体是从可再生胚性细胞悬浮液中分离出来的。以下参数被发现具有高度影响力:电穿孔缓冲液、聚乙二醇处理及其在电穿孔前的持续时间、使用热休克和嵌合基因构建体。瞬时表达 uidA 基因的原位检测显示,DNA 引入的最大频率达到了总原生质体的 1.8%。由于最近已经从香蕉原生质体中高频再生出植物,因此目前的结果可能有助于生产转基因香蕉。