The Otto Warburg Center for Biotechnology in Agriculture, Faculty of Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel.
Plant Mol Biol. 1988 May;10(3):185-91. doi: 10.1007/BF00027395.
The expression plasmid vector pUC8CaMVCAT, containing the chloramphenicol acetyl transferase (CAT) gene, was encapsulated in large unilamellar vesicles (LUV) and introduced into tobacco protoplasts derived from either cell suspension culture or leaf mesophyll. Treatment with liposomes took place in a buffer containing either NaCl or CaCl2, but no polyethylene glycol. The presence of polylysine in the incubation buffer increased the adsorption of liposomes to protoplasts but decreased the efficiency of CAT gene expression.The expression of the introduced CAT gene could be monitored for at least seven days, following the treatment (about 25% acetylation at day 3 as well as at day 7). Plasmid DNA sequences could be detected, apparently unmodified, for at least nine days in the plant cells, though unintegrated in the host genome.
表达质粒载体 pUC8CaMVCAT 含有氯霉素乙酰转移酶(CAT)基因,被包裹在大单层囊泡(LUV)中,并被导入来自悬浮细胞培养或叶肉的烟草原生质体。用含有 NaCl 或 CaCl2 但不含聚乙二醇的缓冲液处理脂质体。在孵育缓冲液中添加聚赖氨酸会增加脂质体对原生质体的吸附,但会降低 CAT 基因表达的效率。在处理后(第 3 天和第 7 天约有 25%的乙酰化),至少可以监测到导入的 CAT 基因表达至少 7 天。尽管未整合到宿主基因组中,但在植物细胞中,质粒 DNA 序列至少可在 9 天内被检测到,且未被修饰。