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GrainGenes Reference Report: PRT-12-671

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Reference	PRT-12-671
Title	Gene transfer by electroporation into intact scutellum cells of wheat embryos
Journal	Plant Cell Reports
Year	1993
Volume	12
Pages	671-675
Author	Kloti A [ Show all 6 ]
Abstract	Gene transfer into intact cells was achieved by electroporating zygotic wheat embryos without any special pretreatment Electroporation was tissue specific in so far as scutellum cells were found to be much more susceptible to gene transfer than other cell types of the embryo The orientation of the embryos in the electroporation chamber also influenced die number of transformed scutellum cells; during electroporation, as in electrophoresis, the negatively charged plasmid DNA molecules seemed to move towards the positive electrode Therefore, the embryos were arranged so that the scutella faced the negative electrode The use of plasmids carrying either two chimeric anthocyanin regulatory genes or a chimeric gusA gene allowed clear identification of transformed cells in the scutellum. On some of the embryos, more than 100 transformed scutellum cells were found after electroporation with single electric pulses of 275 V/cm discharged from a 960-microfarad capacitor and with 100 microgram DNA/ml electroporation buffer. Using the anthocyanin marker system, visibly transformed cells grew to produce red sectors.
Keyword	anthocyanins [ Show all 14 ]

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