Ballas N, Zakai N, Loyter A
Exp Cell Res. 1987 May;170(1):228-34. doi: 10.1016/0014-4827(87)90132-7.
Petunia and carrot protoplasts have been transformed with the plasmid pCaMVCAT by the use of polyethyleneglycol (PEG) as a facilitator. Transformation was revealed by the appearance of the chloramphenicol-acetyl transferase (CAT) enzyme within the transformed cells. Maximal activity of the CAT enzyme was detected within 15 h following transformation, while after 60 h, its activity was significantly reduced, indicating transient expression of the CAT gene. The efficiency of transformation was highly dependent on the presence of CaCl2 in the transformation system, was stimulated by non-functional carrier DNA and was independent on the molecular weight (MW) of PEG used.
矮牵牛和胡萝卜原生质体已通过使用聚乙二醇(PEG)作为促进剂,用质粒pCaMVCAT进行了转化。转化细胞中氯霉素乙酰转移酶(CAT)的出现表明发生了转化。转化后15小时内检测到CAT酶的最大活性,而60小时后,其活性显著降低,表明CAT基因的瞬时表达。转化效率高度依赖于转化体系中氯化钙的存在,受无功能载体DNA的刺激,且与所用PEG的分子量无关。