Department of Plant Breeding, Swedish University of Agricultural Sciences, Box 7003, S-750 07, Uppsala, Sweden.
Plant Cell Rep. 1990 Jul;9(2):105-8. doi: 10.1007/BF00231560.
Transformation of Brassica napus mesophyll protoplasts was performed with the ß-glucuronidase gene fusion system. After electroporation, transient expression in protoplasts transformed directly after isolation was about 1 to 2 per million. By the use of 2,6-dichloro-benzonitrile, a non-toxic inhibitor of cell wall synthesis, and in the presence of 5% polyethyleneglycol, transformation of the cell material was performed three days after isolation. At that time, about 25-30% of the protoplasts had reached the first S-phase of the mitotic cycle. A 1000 fold increase of protoplasts expressing the ß-glucoronisidase gene transiently was obtained, in the partly synchronized protoplasts, compared to those transformed directly after isolation.
油菜叶片原生质体的转化采用β-葡萄糖醛酸酶基因融合系统。电穿孔后，直接分离转化的原生质体中的瞬时表达约为每百万分之一的 1 到 2。通过使用 2,6-二氯苯腈，一种非毒性的细胞壁合成抑制剂，以及在 5%聚乙二醇的存在下，在分离后三天进行细胞材料的转化。此时，约 25-30%的原生质体已经达到有丝分裂周期的第一个 S 期。与直接分离转化的原生质体相比，在部分同步化的原生质体中，瞬时表达β-葡萄糖醛酸酶基因的原生质体增加了 1000 倍。
