Lipofectin: direct gene transfer to higher plants using cationic liposomes
Lipofectin: direct gene transfer to higher plants using cationic liposomes
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Lipofectin：使用阳离子脂质体将基因直接转移至高等植物
DOI：
10.1007/bf00229218
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发表时间：
1991
影响因子：
5.4
通讯作者：
H. Koop
中科院分区：
文献类型：
--
作者：
B. Spörlein;H. Koop
概述最近已经表明，脂质体，一种市售的阳离子脂质体制剂，能够转染动物和植物细胞系。在这里，它的毒性进行了分析，高等植物原生质体和用于瞬时表达和稳定的转化实验与叶肉原生质体的烟草和烟草plumbaginifolia。在CaMV-35 S启动子控制下的β-葡萄糖醛酸酶基因（GUS）的瞬时表达低于通过聚乙二醇引入相同基因后的瞬时表达。通过转移新霉素磷酸转移酶基因（NPTII）和随后的培养和再生的选择下，卡那霉素，稳定转化的植物回收后，使用Lipofectin在各种协议，有或没有额外的应用电穿孔。稳定转化的效果与用PEG和/或电穿孔实现的效果相当。转化体的确认包括测定基因产物的酶活性、基因组印迹和抗性表型转移至由自交的初级转化体产生的后代。
SummaryIt 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.
DOI：
10.1073/pnas.84.21.7413
发表时间：
1987-11-01
影响因子：
11.1
作者：
FELGNER, PL;GADEK, TR;DANIELSEN, M
通讯作者：
DANIELSEN, M