Citation
Che Radziah C. M. Z., . and Marziah M., . and Maheran Abd. Aziz, . and Siti Khalijah Daud, . (2000) Using GFP gene in optimization of genetic transformation protocols in Musa spp. cv. Berangan. [Proceedings Paper]
Abstract
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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Abstract
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.
Additional Metadata
| Item Type: | Proceedings Paper |
|---|---|
| Additional Information: | Available at Perpustakaan Sultan Abdul Samad Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia. QK710.2 M239 2000 Call Number |
| AGROVOC Term: | Musa sapientum |
| AGROVOC Term: | Bananas |
| AGROVOC Term: | Musaceae |
| AGROVOC Term: | Genetic transformation |
| AGROVOC Term: | Proteins |
| AGROVOC Term: | Genetic markers |
| AGROVOC Term: | Callus |
| AGROVOC Term: | Cells |
| AGROVOC Term: | Gene expression |
| AGROVOC Term: | Plant establishment |
| Geographical Term: | MALAYSIA |
| Depositing User: | Ms. Suzila Mohamad Kasim |
| Last Modified: | 24 Apr 2025 05:15 |
| URI: | http://webagris.upm.edu.my/id/eprint/12940 |
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