United States Department of Agriculture, Science and Education Administration, Agricultural Research, Plant Physiology Institute, Beltsville, Maryland 20705.
Plant Physiol. 1979 Apr;63(4):683-6. doi: 10.1104/pp.63.4.683.
Protoplasts prepared from cultured tobacco cells were treated with ColE1-kan plasmid DNA, a hybrid of ColE1 and pSC105 plasmids bearing a gene for kanamycin resistance. The conditions employed permitted the uptake or irreversible binding of 2.9% of the added DNA in acid-insoluble form. Upon commencement of division, the treated cells were plated in agar medium containing kanamycin and differentiating hormones. Plantlets or shoots obtained as presumptive transformants were further tested on kanamycin medium by subculturing small leaf pieces. No evidence was obtained for expression of the kanamycin resistance gene of ColE1-kan in tobacco tissue.
从培养的烟草细胞中制备的原生质体用 ColE1-kan 质粒 DNA 处理，ColE1-kan 是 ColE1 和 pSC105 质粒的杂交体，带有卡那霉素抗性基因。所采用的条件允许以酸不溶性形式摄取或不可逆结合添加 DNA 的 2.9%。在开始分裂时，将处理过的细胞铺在含有卡那霉素和分化激素的琼脂培养基上。作为假定转化体获得的植物或芽进一步通过在卡那霉素培养基上进行小叶片的继代培养进行测试。未在烟草组织中观察到 ColE1-kan 的卡那霉素抗性基因的表达证据。
