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

• Autorzy: Lai K.-S. , Abdullah P. , Yusoff K. , Mahmood M.
• Języki publikacji: EN

Abstrakty

EN

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 postbombardment 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.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2011
• Tom: 33
• Numer: 6
• Opis fizyczny: p.2547-2552,fig.,ref.

Twórcy

autor

Lai K.-S.

• Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia

autor

Abdullah P.

• Department of Cell and Molecular Biology, Faculty of Biotechnologyand Biomolecular Sciences, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia
• Institute of Bioscence, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia

autor

Yusoff K.

• Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia
• Institute of Bioscence, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia

autor

Mahmood M.

• Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia
• Institute of Bioscence, Universiti Putra Malaysia (UPM), 43400 Serdang, Selangor DE, Malaysia

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