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  • Title: [Developing transgenic barley lines producing human lactoferrin using mutant alfa-tubulin gene as selective marker gene].
    Author: Tanasienko IV, Emets AI, Pirko IaV, Korkhovoĭ VI, Abumkhadi N, Blium IaB.
    Journal: Tsitol Genet; 2011; 45(1):3-10. PubMed ID: 21446153.
    Abstract:
    Method of biolistic transformation was used for genetic improvement of commercial barley cultivars (Oksamitoviy, Vodogray and Getman). The plasmid pHLFTuBA was used for particle bombardment that consists of the hLF gene under the control of the barley glutelin B-1 promoter and a selectable marker gene, alpha-tubulin conferring resistance to trifluralin (dinitroanilinr herbicide). Preliminary screening of different trifluralin concentration range from 0,1 to 30 microM was tested for determination of effective selective agent concentration. Two transgenic barley line of genotype Oksamitiviy and transgenic callus line of cultivar Getman were obtained after selection on 10 microM of trifluralin. To confirm the transgenic nature of regenerated plants, the PCR analysis was carried out. The 734bp length fragment of hLFgene was amplified from both regenerated plants.
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