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- W1013711245 abstract "Kulonboző in vitro rendszerekben kimutattuk az agrobakterium hatekony fizikai kapcsolodasat es vir indukciojat. Husz kenyerbuza fajta novenyregeneracios kepessegenek osszehasonlito elemzeseből (portokkultura, eretlen es erett embrio, erett szkutellum) megallapitottuk, hogy a leghatekonyabb rendszer az erett embrion alapszik, de univerzalisan kiemelkedő fajtat nem talaltunk. Az erett embrio agrobakteriumos transzformaciojanak egyes parametereinek optimalizaciojaval 45% tranziens genexpressziot ertunk el. Vegul gyors es hatekony in planta transzformacios modszert fejlesztettunk ki buza csiranovenyekre, melyet kiterjesztettunk mas gabonafelekre is. Az in planta transzformacioval kapott transzgenikus esemenyek gyors kimutatasanak erdekeben ket, a pCAMBIA1391z jelzesű alapvektorbol keszitett expresszios vektort ugy modositottunk, hogy kozos szelekcios markergenjuk (hptII) buzaban nem eleg aktiv (35S) promoteret egy erős konstitutiv promoterrel (Act1) csereltuk fel (pKOL1). Ezutan a riporter genekhez (intront tartalmazo gfp vagy gusA) kapcsoltunk egy masik konstitutiv (Ubi1) promotert (pKOL2 illetve pKOL3), majd ezt felvaltottuk egy endospermium-specifikus promoterrel (pKOL4 ill. pKOL5). Vegul keszitettunk ket olyan vektort is, melyek vagy egy nagy molekulatomegű (HMW) glutenin alegysegfeherje genjet (1Ax2*B, pKOL6), vagy pedig a ‘Porcine Epidemic Diarrhea’ virus egy antigen feherje genjet (pKOL7) tartalmaztak egyarant az emlitett endospermium-specifikus promoterhez kapcsolva. | In the early steps of fast in planta transformation we have shown that in several culture systems attachment and vir induction of agrobacteria is efficiently completed. Comparative analysis of plant regeneration capacity (anther culture, immature and mature embryo, mature scutellum) in twenty bread wheat cultivars revealed that the mature embryo system is the most efficient, but a universally excellent cultivar was not identified. By the optimization of several parameters for Agrobacterium transformation of mature embryos were optimized a transient gene expression frequency of 45% was reached. Finally, a fast in planta transformation tool was developed on the basis of wheat seedlings, which was extended to other cereals. For the quick detection of transgenic events obtained by in planta transformation two expression vectors (based on pCAMBIA1391z) were modified so that the less active 35S promoter of their selectable marker gene (nptII) was replaced by the strong constitutive Act1 promoter, resulting pKOL1. Next, a constitutive promoter (Ubi1) was fused to each of the two riporter genes (intron containing gfp or gusA, pKOL2 and pKOL3, respectively), which was then exchanged with an endosperm-specific promoter (pKOL4 and pKOL5, resp.). Finally, two more vectors were constructed for the expression of either a HMW glutenin gene (1Ax2*B, pKOL6) or an antigenic protein gene of the Porcine Epidemic Diarrhea virus (pKOL7), both genes controlled by the same endosperm-specific promoter." @default.
- W1013711245 created "2016-06-24" @default.
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- W1013711245 date "2011-01-01" @default.
- W1013711245 modified "2023-09-26" @default.
- W1013711245 title "Hatékony búzatranszformációs módszer kidolgozása = Development of an efficient wheat transformation method" @default.
- W1013711245 hasPublicationYear "2011" @default.
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