BINDING OF COLE1-KAN PLASMID DNA BY TOBACCO PROTOPLASTS - NON-EXPRESSION OF PLASMID GENE

Protoplasts prepared from cultured tobacco [Nicotiana tabacum L. cv. Bright Yellow] 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 of irreversible binding of 2.9% of the added DNA in acid-soluble 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.