Expression and Inheritance of GUS Gene in Transgenic Tobacco Plants

Authors: TİJEN TALAS OĞRAŞ, NERMİN GÖZÜKIRMIZI

Abstract: Agrobacterium tumefaciens-mediated transformation of tobacco leaf discs was performed with plant transformation vec-tors of pBI 121 and pGus-Int. Both carry the neomycin phosphotransferase II ( npt II) gene and, unlike the pBI 121, pGus-Int car-ries a portable plant intron inside the beta-glucuronidase (GUS) gene. In addition to the integration of the marker genes into the genomes of primary transformants, R1 and R2 progenies were confirmed by polymerase chain reaction (PCR) and Southern blot analyses. Histochemical and fluorometric analyses were also performed to determine the activity of the GUS gene. Segregation of progenies on kanamycin-resistance trait was also shown as evidence for the transformation. The analyses showed that the trans-genes were transmitted to subsequent generations in a Mendelian manner and intron sequences enhanced the expression of the GUS gene by 25-30% in some transformants.

Keywords: Agrobacterium tumefaciens,glucuronidase (GUS), intron

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