Using GFP gene in optimization of genetic transformation protocols in Musa spp. (cv. Berangan)
2000
Che Radziah C. M. Z. | Marziah M. | Maheran Abd. Aziz | Siti Khalijah Daud
Green-fluorescent protein (GFP) has been used as a reporter gene in early detection of plant transformation for establishment and optimization of plant transformation protocols. The prime advantage by using GFP is that no exogenously supplied substrate or cofactors need to be applied for fluorescence as in gus assay. A study was conducted in order to assess the effect of this visual marker gene on embryogenic callus viability and the stability of the transformation. We transformed embryogenic callus with green fluorescent protein (GFP) gene using biolistic gun and the helium pressure used are 450, 900, 1100 or 1350 psi with target distance 6, 9 or 12 cm. The cells were then visually examined under microscope every 48 hours to observe the transformed cells. The results shown that the green fluorescent spots could be detected after 48 hours in callus tissues bombarded using the following parameters 450psi, 6cm (36 spots) ; 900psi, 6cm (20 spots) ; I100psi, 9cm (12 spots) and 1350psi, 6cm (28 spots). However, the gene expressions disappeared slowly and vanished after one month. These results indicated that the gene transformations are unstable and modifications were needed to obtain improved and stable transformation.
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تم تزويد هذا السجل من قبل Universiti Putra Malaysia