High frequency of single-copy T-DNA transformants produced after floral dip in CRE-expressing Arabidopsis plants

Transgenic plants that harbor a single copy of the introduced transgene are preferable to those with multiple transgene copies because multiple T-DNA copies correlate with expression variability and susceptibility to silencing. Especially after the commonly used fl oral-dip Agrobacterium -mediated transformation method, the frequency of single-copy transformants is low. The CRE/ loxP recombinase-based strategy to resolve complex T-DNA loci has proven to be successful to effi ciently obtain single-copy T-DNA transformants by directly transforming loxP -containing T-DNA vectors in CRE -expressing Arabidopsis thaliana plants. This chapter describes in detail how to transform three available loxP -containing T-DNA vectors into CRE - producing Arabidopsis C24 plants and subsequently how to analyze the transgenic plants for the T-DNA locus structure.

ISBN:9781617795589

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