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Genetics Section

Yemets, Alla I. [1], Bayer, G. [1], Radchuk, V.V. [2], Bayer, O. [2], Blume, Yaroslav B. [3], Atanassov, A. [4], Baird, Wm. Vance [5].

The development of transformation vectors based upon modified plant tubulin genes as the selectable marker.

A plant transformation and selection system is being developed that utilizes modified tubulin genes (naturally occurring and induced) as a selectable marker in the vector. As such, it is based exclusively on genetic information of plant origin, and represents an alternative to traditional antibiotic resistance gene-based selection systems. The new selection scheme is based upon mechanisms of resistance to anti-mitotic herbicides (i.e., dinitroanilines, DNH, and phosphorothioamidates, PTH) conferred by modified plant microtubules. The transformation/selection method involves the production of altered tubulin isotypes that result from a mutation(s) in the herbicide-binding site. The new system is derived from information acquired from studies: i) characterizing mutant alpha-tubulins from natural biotypes of Eleusine indica that confer resistance to DNH; ii) of Nicotiana plumbaginifolia mutants and inter-specific hybrids resistant to DNH and PTH, and characterization of altered beta-tubulins; and iii) three-dimensional reconstruction of the interactive sites on alpha- and beta-tubulin (both sensitive and resistant molecules) for DNH and PTH, and predication of new sites for such compatible and incompatible interactions. Current efforts focuses on: (a) Production of a selection marker cassette(s) with sequences coding for alpha-/beta-tubulin conferring resistance to DNH and PTH; and (b) Transformation of model plants (e.g., goosegrass and barley) with these experimental constructs, and establishment of the transformation lines through in vitro selection based on survival following exposure to DNH/PTH.


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1 - University of Kiev, Genomics and Biotechnology, Kiev, , Ukraine
2 - Natl. Acad. of Sciences of Ukraine, Institute of Cell Biology and Genetic Engineering, Kiev, , Ukraine
3 - National Academy of Sciences of Ukraine, Institute of Cell Biology and Genetic Engineering, acad. Zabolotnogo str., 148, Kiev, 03143, Ukraine
4 - AgroBioInstitute, Center of Excellence in Plant Biotechnology, 8 Dragan Tzankov Blvd., Sofia, 1164, Bulgaria
5 - Clemson University, Horticulture Department, Poole Agr Cntr, Box 340319, Clemson, South Carolina, 29634-0319, USA

Keywords:
alpha-tubulin
beta-tubulin
dinitroaniline
phosphorothioamidate.

Presentation Type: Poster
Session: 33-56
Location: Salon C, D & E - Gov Ballroom/Hilton
Date: Tuesday, August 16th, 2005
Time: 12:30 PM
Abstract ID:458


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