Abstract
We describe here protocols for isolating genes in maize using Dissociation (Ds) transposons marked with a green fluorescent protein (GFP) transgene. The introduced marker enables the phenotypic scoring of the nonautonomous element and the anchoring of unique primers on the element to facilitate the isolation of the adjacent DNA by PCR. Transposons such as Ds transpose preferentially to sites closely linked to the Ds-launching platform. Based on this transposition behavior, a genetic resource is being created to mobilize a modified Ds element from different starting sites in the genome. Enough transgenic lines are being generated to cover most of the maize genome, allowing the targeted tagging of most genes from a Ds-launching platform located nearby.
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Acknowledgment
We thank Jun Huang and Limei He for helpful suggestions. This work is supported by National Science Foundation Plant Genome Program project DBI-0929350.
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Li, Y., Segal, G., Wang, Q., Dooner, H.K. (2013). Gene Tagging with Engineered Ds Elements in Maize. In: Peterson, T. (eds) Plant Transposable Elements. Methods in Molecular Biology, vol 1057. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-568-2_6
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DOI: https://doi.org/10.1007/978-1-62703-568-2_6
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