Isolation of active DNA-binding nuclear proteins from tomato galls induced by root-knot nematodes |
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| Authors: | Carolina Escobar Fabio Aristizéabal Alfonso Navas Francisca F Del Campo Carmen Fenoll |
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| Institution: | (1) Facultad de Ciencias del Medio Ambiente, Universidad de Castilla-La Mancha, Campus de la Féabrica de Armas, 45071 Toledo, Spain;(2) Present address: Departamento de Farmacia, Facultad de Ciencias, Universidad Nacional de Colombia, Ciudad Universitaria, A.A. 14490 Bogotéa, Colombia;(3) Departamento de Biologéia, Universidad Autéonoma de Madrid, Cantoblanco, 28049 Madrid, Spain;(4) Departamento de Biodiversidad y Biologéia Evolutiva, Museo Nacional de Ciencias Naturales, CSIC, Josée Gutiéerrez, Abascal 2, 28006 Madrid |
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| 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.
Both authors have contributed equally to this work. |
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| Keywords: | galls gel retardation nematode nuclear protein promoter |
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