The Mutated Acetolactate Synthase Gene from Rice as a Non-Antibiotic Selection Marker for Transformation of Bamboo CellsReportar como inadecuado




The Mutated Acetolactate Synthase Gene from Rice as a Non-Antibiotic Selection Marker for Transformation of Bamboo Cells - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Previously, we developed a particle bombardment-mediated transformation protocol in Phyllostachys nigra bamboo by expressing hygromycin phosphotransferase gene HPT and neomycin phosphotransferase II gene NPT II. Although these marker genes could introduce to several tissue cultured organs e.g. leaves, buds, and calli of Phyllostachs bamboo species, some organs showed a high susceptibility and-or a low selectivity to hygromycin and kanamycin. In this report, therefore, we describe advantages and technical details for generating stable transgenic bamboo cells using the particle bombardment method with the mutated-acetolactate synthase gene mALS from rice W548L-S627IOsALS as a non-antibiotic selection marker. A facile and efficient transformation was achieved with the mALS gene and enhanced fluorescent protein gene mCherry. Approximately 490 and 1400 mCherry-expressing cells-dish-shot in average were observed in both P. bambusoides and P. nigra under fluorescent stereo-microscope. Stable transgenic bamboo cell lines were generated in a selection medium supplemented with 0.1 μM of bispyribac-sodium BS as ALS inhibitor. The integration of mALS gene was identified by in vivo ALS enzyme assay and a PCR-restriction fragment length polymerphism RFLP based detection procedures.

KEYWORDS

Bamboo; Mutated Acetolactate Synthase Gene; Particle Bombardment; Suspension Culture

Cite this paper

S. Ogita, N. Kikuchi, T. Nomura and Y. Kato -The Mutated Acetolactate Synthase Gene from Rice as a Non-Antibiotic Selection Marker for Transformation of Bamboo Cells,- American Journal of Plant Sciences, Vol. 3 No. 3, 2012, pp. 368-372. doi: 10.4236-ajps.2012.33044.





Autor: Shinjiro Ogita, Nanaka Kikuchi, Taiji Nomura, Yasuo Kato

Fuente: http://www.scirp.org/



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