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Efficient enrichment cloning of TAL effector genes from Xanthomonas

Many plant-pathogenic xanthomonads use a type III secretion system to translocate Transcription Activator-Like (TAL) effectors into eukaryotic host cells where they act as transcription factors. Target genes are induced upon binding of a TAL effector to double-stranded DNA in a sequence-specific man...

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Autores principales: Tran, T.T., Doucouré, H., Hutin, M., Jaimes Niño, L.M., Szurek, B., Cunnac, S., Koebnik, R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6138780/
https://www.ncbi.nlm.nih.gov/pubmed/30225203
http://dx.doi.org/10.1016/j.mex.2018.08.014
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author Tran, T.T.
Doucouré, H.
Hutin, M.
Jaimes Niño, L.M.
Szurek, B.
Cunnac, S.
Koebnik, R.
author_facet Tran, T.T.
Doucouré, H.
Hutin, M.
Jaimes Niño, L.M.
Szurek, B.
Cunnac, S.
Koebnik, R.
author_sort Tran, T.T.
collection PubMed
description Many plant-pathogenic xanthomonads use a type III secretion system to translocate Transcription Activator-Like (TAL) effectors into eukaryotic host cells where they act as transcription factors. Target genes are induced upon binding of a TAL effector to double-stranded DNA in a sequence-specific manner. DNA binding is governed by a highly repetitive protein domain, which consists of an array of nearly identical repeats of ca. 102 base pairs. Many species and pathovars of Xanthomonas, including pathogens of rice, cereals, cassava, citrus and cotton, encode multiple TAL effectors in their genomes. Some of the TAL effectors have been shown to act as key pathogenicity factors, which induce the expression of susceptibility genes to the benefit of the pathogen. However, due to the repetitive character and the presence of multiple gene copies, high-throughput cloning of TAL effector genes remains a challenge. In order to isolate complete TAL effector gene repertoires, we developed an enrichment cloning strategy based on • genome-informed in silico optimization of restriction digestions, • selective restriction digestion of genomic DNA, and • size fractionation of DNA fragments. Our rapid, cheap and powerful method allows efficient cloning of TAL effector genes from xanthomonads, as demonstrated for two rice-pathogenic strains of Xanthomonas oryzae from Africa.
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spelling pubmed-61387802018-09-17 Efficient enrichment cloning of TAL effector genes from Xanthomonas Tran, T.T. Doucouré, H. Hutin, M. Jaimes Niño, L.M. Szurek, B. Cunnac, S. Koebnik, R. MethodsX Agricultural and Biological Science Many plant-pathogenic xanthomonads use a type III secretion system to translocate Transcription Activator-Like (TAL) effectors into eukaryotic host cells where they act as transcription factors. Target genes are induced upon binding of a TAL effector to double-stranded DNA in a sequence-specific manner. DNA binding is governed by a highly repetitive protein domain, which consists of an array of nearly identical repeats of ca. 102 base pairs. Many species and pathovars of Xanthomonas, including pathogens of rice, cereals, cassava, citrus and cotton, encode multiple TAL effectors in their genomes. Some of the TAL effectors have been shown to act as key pathogenicity factors, which induce the expression of susceptibility genes to the benefit of the pathogen. However, due to the repetitive character and the presence of multiple gene copies, high-throughput cloning of TAL effector genes remains a challenge. In order to isolate complete TAL effector gene repertoires, we developed an enrichment cloning strategy based on • genome-informed in silico optimization of restriction digestions, • selective restriction digestion of genomic DNA, and • size fractionation of DNA fragments. Our rapid, cheap and powerful method allows efficient cloning of TAL effector genes from xanthomonads, as demonstrated for two rice-pathogenic strains of Xanthomonas oryzae from Africa. Elsevier 2018-09-04 /pmc/articles/PMC6138780/ /pubmed/30225203 http://dx.doi.org/10.1016/j.mex.2018.08.014 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Agricultural and Biological Science
Tran, T.T.
Doucouré, H.
Hutin, M.
Jaimes Niño, L.M.
Szurek, B.
Cunnac, S.
Koebnik, R.
Efficient enrichment cloning of TAL effector genes from Xanthomonas
title Efficient enrichment cloning of TAL effector genes from Xanthomonas
title_full Efficient enrichment cloning of TAL effector genes from Xanthomonas
title_fullStr Efficient enrichment cloning of TAL effector genes from Xanthomonas
title_full_unstemmed Efficient enrichment cloning of TAL effector genes from Xanthomonas
title_short Efficient enrichment cloning of TAL effector genes from Xanthomonas
title_sort efficient enrichment cloning of tal effector genes from xanthomonas
topic Agricultural and Biological Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6138780/
https://www.ncbi.nlm.nih.gov/pubmed/30225203
http://dx.doi.org/10.1016/j.mex.2018.08.014
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