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Factors influencing transient gene expression in electroporated tall fescue (Festuca arundinacea Schreb.) protoplasts

For the rapid establishment of optimal conditions for a genetic transformation system for tall fescue, several factors influencing transient gene expression were studied in protoplasts, after the reporter β-glucuronidase (GUS) gene was introduced by electroporation. In a time-course study of transient gene expression, GUS activity peaked at 24 h after electroporation. Among the different field strength conditions tested, maximum GUS activity was observed at 750 V/cm. Increases in the amount of plasmid DNA to 80 μg/ml led to increased GUS activity. GUS activities increased in linear fashion with increasing protoplast densities up to 2 x 10⁶/ml. Age of suspension cells from which protoplasts were derived influenced transient expression with maximum GUS activity obtained in 3 and 5 day old suspensions. These results show that transient expression studies can be used to optimize electroporation parameters rapidly. Results of such rapid assays can be used as a basis for further studies on stable transformation of this important turf-grass species. / Master of Science

Identiferoai:union.ndltd.org:VTETD/oai:vtechworks.lib.vt.edu:10919/44549
Date05 September 2009
CreatorsPenmetsa, Ramachandra V.
ContributorsCrop and Soil Environmental Sciences
PublisherVirginia Tech
Source SetsVirginia Tech Theses and Dissertation
LanguageEnglish
Detected LanguageEnglish
TypeThesis, Text
Formatviii, 55 leaves, BTD, application/pdf, application/pdf
RightsIn Copyright, http://rightsstatements.org/vocab/InC/1.0/
RelationOCLC# 28315749, LD5655.V855_1992.P466.pdf

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