Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system
Genetic transformation system of Dendrobium Sonia 17 was optimized using green fluorescent protein (GFP) and β-glucuronidase (GUS) gene as the reporter systems. The 35S-sgfp-TYG-nos (p35S) and pSMDFR, carrying sgfp and gusA gene, respectively, were co-bombarded into the calluses. Parameters optimize...
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| Format: | Article |
| Language: | English English |
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Springer
2005
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| Online Access: | http://psasir.upm.edu.my/id/eprint/18372/ http://psasir.upm.edu.my/id/eprint/18372/1/Optimization%20of%20biolistic%20bombardment%20parameters%20for%20Dendrobium%20sonia%2017%20calluses%20using%20GFP%20and%20GUS%20as%20the%20reporter%20system.pdf |
| _version_ | 1848843493643911168 |
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| author | Tee, Chong Siang Maziah, Mahmood |
| author_facet | Tee, Chong Siang Maziah, Mahmood |
| author_sort | Tee, Chong Siang |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Genetic transformation system of Dendrobium Sonia 17 was optimized using green fluorescent protein (GFP) and β-glucuronidase (GUS) gene as the reporter systems. The 35S-sgfp-TYG-nos (p35S) and pSMDFR, carrying sgfp and gusA gene, respectively, were co-bombarded into the calluses. Parameters optimized were acceleration pressure, target distance, gold particle size, pre-bombardment cultured time, plasmid DNA precipitation, total plasmid DNA and the ratio of the plasmids co-bombarded. Both reporter systems responded similarly to the bombardment parameters investigated. Based on the GUS/GFP spot counts, the GFP expression rate was higher than that for GUS under the control of the same promoter, CaMV 35S. GFP could be used as the reporter system for the co-bombardment as it was rapid and non-destructive system to monitor the transformed tissues. A combination of GFP and antibiotic resistance gene was used to select stable putative transformants. © Springer 2005. |
| first_indexed | 2025-11-15T08:15:54Z |
| format | Article |
| id | upm-18372 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:15:54Z |
| publishDate | 2005 |
| publisher | Springer |
| recordtype | eprints |
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| spelling | upm-183722016-01-21T03:40:54Z http://psasir.upm.edu.my/id/eprint/18372/ Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system Tee, Chong Siang Maziah, Mahmood Genetic transformation system of Dendrobium Sonia 17 was optimized using green fluorescent protein (GFP) and β-glucuronidase (GUS) gene as the reporter systems. The 35S-sgfp-TYG-nos (p35S) and pSMDFR, carrying sgfp and gusA gene, respectively, were co-bombarded into the calluses. Parameters optimized were acceleration pressure, target distance, gold particle size, pre-bombardment cultured time, plasmid DNA precipitation, total plasmid DNA and the ratio of the plasmids co-bombarded. Both reporter systems responded similarly to the bombardment parameters investigated. Based on the GUS/GFP spot counts, the GFP expression rate was higher than that for GUS under the control of the same promoter, CaMV 35S. GFP could be used as the reporter system for the co-bombardment as it was rapid and non-destructive system to monitor the transformed tissues. A combination of GFP and antibiotic resistance gene was used to select stable putative transformants. © Springer 2005. Springer 2005-01 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/18372/1/Optimization%20of%20biolistic%20bombardment%20parameters%20for%20Dendrobium%20sonia%2017%20calluses%20using%20GFP%20and%20GUS%20as%20the%20reporter%20system.pdf Tee, Chong Siang and Maziah, Mahmood (2005) Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system. Plant Cell, Tissue and Organ Culture, 80 (1). pp. 77-89. ISSN 0167-6857 http://dx.doi.org/10.1007/s11240-004-9144-6 Plant genetic transformation Dendrobium - Genetics Green fluorescent protein doi:10.1007/s11240-004-9144-6 English |
| spellingShingle | Plant genetic transformation Dendrobium - Genetics Green fluorescent protein Tee, Chong Siang Maziah, Mahmood Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system |
| title | Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system |
| title_full | Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system |
| title_fullStr | Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system |
| title_full_unstemmed | Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system |
| title_short | Optimization of biolistic bombardment parameters for Dendrobium sonia 17 calluses using GFP and GUS as the reporter system |
| title_sort | optimization of biolistic bombardment parameters for dendrobium sonia 17 calluses using gfp and gus as the reporter system |
| topic | Plant genetic transformation Dendrobium - Genetics Green fluorescent protein |
| url | http://psasir.upm.edu.my/id/eprint/18372/ http://psasir.upm.edu.my/id/eprint/18372/ http://psasir.upm.edu.my/id/eprint/18372/ http://psasir.upm.edu.my/id/eprint/18372/1/Optimization%20of%20biolistic%20bombardment%20parameters%20for%20Dendrobium%20sonia%2017%20calluses%20using%20GFP%20and%20GUS%20as%20the%20reporter%20system.pdf |