Turkish Journal of Botany

Construction of Vectors for npt-II Gene Transfer Into Tobacco and the Effects of Different Antibiotics on Morphogenesis




The intermediate vectors pAZ1neol and pAZ2neol, containing the neomycinphosphotransferase II gene (npt-II) under the control of single or double promoters of gene 5 of T-DNA pTiAch5, were constructed. The agrobacterial strain containing stable Kmr cointegrate was obtained by crossing three parental strains, i.e. HB101 (containing kanamycin resistant (Kmr) plasmid), GJ23 (plasmids- R64drd11 and pGJ28) and GV3101 (pGV3850). Transformation of tobacco plant cells was carried out by co-cultivation with A. tumefaciens, and an antibiotic ampiox capable of suppressing A. tumefaciens growth with no effect on morphogenesis processes was chosen. Kanamycin-resistant regenerants were obtained from tobacco leaf disks. It was found that the expression level of the chimaeric nptII gene directed by single or double promoters of gene 5 of T-DNA pTiAch5 did not differ in transformed plant cells at the level of electrophoretic analysis.


Agrobacterium tumefaciens, Ti-plasmid, T-DNA, Transformation

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