Monohybrid and dihybrid segregations in the progenies of tobacco transformed for kanamycin resistance with a Ti-vector system |
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Authors: | Patricia Viegas Helena Mathews C R Bhatia N K Notani |
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Institution: | 1. Molecular Biology and Agriculture Division, Bhabha Atomic Research Centre, 400 085, Trombay, Bombay, India 2. Bio-Organic Division, Bhabha Atomic Research Centre, 400 085, Trombay, Bombay, India 3. Nuclear Agriculture Division, Bhabha Atomic Research Centre, 400 085, Trombay, Bombay, India
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Abstract: | A chimeric DNA construction having nopaline synthase promoter, coding sequences of neomycin phosphotransferase gene conferring resistance to antibiotic kanamycin and OCS (octopine synthase) polyadenylation sequences bracketed by T-DNA ends was transferred to tobacco. Leaf discs were infected withA. tumefaciens containing disarmed, cointegrate plasmid pGV3850:: 1103 and allowed to form a callus in the presence of kanamycin. Shoots regenerated from infected leaf discs either through the callus or arising directly were further selected for their ability to root in kanamycin-containing media. Among the nine transgenic plants that were progeny tested, the transferred bacterial gene segregated as monohybrid ratio (3 KanR: 1 Kans) in seven. Segregation data of two plant progenies indicated the presence of two independent loci of KanR DNA insertion (15 KanR: 1 Kan s ). Back-cross segregation data were consistent with the monohybrid or independent assortment of duplicate factors. Thus in the two cases, a minimum independent integration of two copies of T-DNA each with a KanR marker is inferred. |
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