Authors: S Komarnytsky A Gaume A Garvey N Borisjuk I Raskin
Publish Date: 2004/02/10
Volume: 22, Issue: 10, Pages: 765-773
Abstract
Requirement for antibioticresistance selection markers and difficulty in identifying transgenes with the highest expression levels remain the major obstacles for rapid production of recombinant proteins in plants An alternative approach to producing transgenic plants free of antibioticresistance markers is the phenotypicbased selection with rootproliferation genes rol genes of Agrobacterium rhizogenes By using Agrobacterium tumefaciens harboring the pRYG transformation vector with a cluster of rol genes linked to a heterologous gene of interest we have developed a rapid transformation tool using hairy root formation as a selection marker The expression of βglucuronidase in newly induced transgenic tobacco roots could be detected as early as 12 days after inoculation Higher levels of transgene expression in the roots correlated positively with the rates of root elongation on hormonefree medium and thus could be used for positive selection When tobacco plants were transformed with pRYG harboring the expression cassette for secreted alkaline phosphatase SEAP the release of SEAP from roots of the fully regenerated transgenic plants could be quantified at rates as high as 28 μg/g root dry weight per dayThe authors thank Dr Stanton Gelvin Purdue University for the mas2′ promoter and Dr Thomas Schmulling Freie Universitat Berlin for the cluster of rol genes We are grateful to Ivan Jenkins for his technical assistance in the greenhouse and to Nir Yakoby for helpful discussions and for reviewing the manuscript A Gaume is a recipient of a Swiss Science Foundation postdoctoral fellowship
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