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Transient expression of gus gene in intact seed embryos of Indica rice after electroporation-mediated gene delivery
Authors:Ashok Chaudhury  Satish C. Maheshwari  Akhilesh K. Tyagi
Affiliation:(1) Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, 110021 New Delhi, India
Abstract:Summary Two-day-old germinating intact seed embryos of Oryza sativa variety Basmati 370 were electroporated with a view to examine suitability of this system for gene delivery. The experiments were done with a plasmid having gus gene under the control of CaMV 35S promoter. Spectrofluorophotometric GUS assay revealed high activity of the introduced gene when embryos were given three electrical pulses at 1600 V cm-1 and 100 mgrF capacitance with a pulse length of 75 ms. Additionally, histochemical localization of GUS activity in seedlings and various organs such as leaves, coleoptiles and roots was also done. Expression of GUS activity was studied up to 15 days and found to be organ-specific, thereby showing that embryos can indeed serve as efficient recipient system. Use of cycloheximide revealed that GUS activity appears as a result of early protein synthesis after electroporation and is substantially stable in vivo.
Keywords:electroporation  germinating seed embryo  GUS  Oryza sativa  transient gene expression
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