Abstract
We describe 2 methods for extraction of DNA-binding proteins from root-knot nematode feeding sites (ie, galls). DNA-binding activity was assayed by electrophoretic mobility shift assays using fragments from the root-knot nematode-responsiveLEMMI9 and 35S promoters. In noninfected tissue, the method based on nuclei enrichment through a Percoll cushion was superior for isolation of DNA-protein binding activity with both promoters. With infected roots; the method based on crude extracts performed better with theLEMMI9 promoter, whereas nuclei-enriched extracts worked better with the 35S promoter. Therefore, both methods can be used to extract proteins for DNA-binding assays from infected roots, but the method of choice may depend on the promoter under study.
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Escobar, C., Aristizéabal, F., Navas, A. et al. Isolation of active DNA-binding nuclear proteins from tomato galls induced by root-knot nematodes. Plant Mol Biol Rep 19, 375–376 (2001). https://doi.org/10.1007/BF02772837
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DOI: https://doi.org/10.1007/BF02772837