Botanical Institute of the Ludwig-Maximilian University, Menzinger Str. 67, W-8000, München 19, Germany.
Theor Appl Genet. 1991 Nov;83(1):1-5. doi: 10.1007/BF00229218.
It has recently been shown that lipofectin, a commercially available preparation of cationic liposomes is capable of animal and plant cell line transfection. Here, it is analyzed with respect to its toxicity for higher plant protoplasts and used for transient expression and stable transformation experiments with mesophyll protoplasts of Nicotiana tabacum and Nicotiana plumbaginifolia. Transient expression of the β-glucuronidase gene (GUS) under control of the CaMV-35S-promoter was lower than after introduction of the same gene by polyethylene glycol. By transferring the neomycin phosphotransferase gene (NPTII) and subsequent culture and regeneration under selection with kanamycin, stably transformed plants were recovered after using Lipofectin in various protocols with or without additional application of electroporation. Efficiencies of stable transformation were comparable to those achieved with PEG and/or electroporation. Confirmation of transformants included assaying the enzyme activity of the gene product, genomic blotting, and transfer of the resistant phenotype to the progeny produced from selfed primary transformants.
最近的研究表明，脂质体转染试剂（Lipofectin）是一种商品化的阳离子脂质体，能够对动植物细胞系进行转染。本文对其用于高等植物原生质体的毒性进行了分析，并将其用于烟草和落地生根叶肉原生质体的瞬时表达和稳定转化实验。CaMV-35S 启动子控制下的β-葡萄糖醛酸酶（GUS）基因的瞬时表达低于聚乙二醇介导的相同基因的表达。通过转移新霉素磷酸转移酶基因（NPTII），并在选择培养基上继代培养和再生，用脂质体在不同的方案中进行转化，无论是单独使用还是与电穿孔联合使用，都可以获得稳定转化的植株。稳定转化的效率与 PEG 和/或电穿孔的效率相当。转化体的验证包括测定基因产物的酶活性、基因组印迹和将抗性表型转移到自交初级转化体产生的后代中。
