An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus
In this study, we report an optimization of particle bombardment transformation system for Centella asiatica callus. A total of eight parameters affecting the genetic transformation system were optimized using the synthetic green fluorescent protein (sGFP) as a reporter driven by the CaMV 35S promot...
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| Format: | Article |
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Springer-Verlag
2011
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| author | Lai, Kok Song Abdullah, Puad Yusoff, Khatijah Mahmood, Maziah |
| author_facet | Lai, Kok Song Abdullah, Puad Yusoff, Khatijah Mahmood, Maziah |
| author_sort | Lai, Kok Song |
| collection | UPM |
| description | In this study, we report an optimization of particle bombardment transformation system for Centella asiatica callus. A total of eight parameters affecting the genetic transformation system were optimized using the synthetic green fluorescent protein (sGFP) as a reporter driven by the CaMV 35S promoter. The results indicated that DNA delivery conditions of 9-cm target distance, 1,100 psi helium pressure, 1.0 μm gold particles size, 27 mmHg chamber vacuum pressure, 2 times number of bombardment, spermidine as precipitation agent, 60 h post-bombardment incubation time and 2 μg plasmid DNA concentration were optimal for C. asiatica callus transformation. The expression of sGFP was monitored using fluorescence microscope and further confirmed using RT-PCR. This optimized genetic transformation system is applicable for rapid transient gene analysis and transgenic C. asiatica production. |
| first_indexed | 2024-03-06T07:53:46Z |
| format | Article |
| id | upm.eprints-22377 |
| institution | Universiti Putra Malaysia |
| last_indexed | 2024-03-06T07:53:46Z |
| publishDate | 2011 |
| publisher | Springer-Verlag |
| record_format | dspace |
| spelling | upm.eprints-223772015-12-07T05:02:12Z http://psasir.upm.edu.my/id/eprint/22377/ An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus Lai, Kok Song Abdullah, Puad Yusoff, Khatijah Mahmood, Maziah In this study, we report an optimization of particle bombardment transformation system for Centella asiatica callus. A total of eight parameters affecting the genetic transformation system were optimized using the synthetic green fluorescent protein (sGFP) as a reporter driven by the CaMV 35S promoter. The results indicated that DNA delivery conditions of 9-cm target distance, 1,100 psi helium pressure, 1.0 μm gold particles size, 27 mmHg chamber vacuum pressure, 2 times number of bombardment, spermidine as precipitation agent, 60 h post-bombardment incubation time and 2 μg plasmid DNA concentration were optimal for C. asiatica callus transformation. The expression of sGFP was monitored using fluorescence microscope and further confirmed using RT-PCR. This optimized genetic transformation system is applicable for rapid transient gene analysis and transgenic C. asiatica production. Springer-Verlag 2011-11 Article PeerReviewed Lai, Kok Song and Abdullah, Puad and Yusoff, Khatijah and Mahmood, Maziah (2011) An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus. Acta Physiology Plantarum, 33 (6). pp. 2547-2552. ISSN 0137-5881; ESSN: 1861-1664 http://link.springer.com/article/10.1007%2Fs11738-011-0786-9 10.1007/s11738-011-0786-9 |
| spellingShingle | Lai, Kok Song Abdullah, Puad Yusoff, Khatijah Mahmood, Maziah An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus |
| title | An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus |
| title_full | An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus |
| title_fullStr | An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus |
| title_full_unstemmed | An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus |
| title_short | An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus |
| title_sort | efficient protocol for particle bombardment mediated transformation of centella asiatica callus |
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