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An efficient protocol for particle bombardment-mediated transformation of Centella asiatica callus


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


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.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Institute of Bioscience
DOI Number: https://doi.org/10.1007/s11738-011-0786-9
Publisher: Springer-Verlag
Keywords: Centella asiatica; SGFP expression; Particle bombardment RT-PCR; Transformation
Depositing User: Nurul Ainie Mokhtar
Date Deposited: 07 Dec 2015 05:02
Last Modified: 07 Dec 2015 05:02
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1007/s11738-011-0786-9
URI: http://psasir.upm.edu.my/id/eprint/22377
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