Efficient enrichment cloning of TAL effector genes from Xanthomonas
| Autor: | Ralf Koebnik, Mathilde Hutin, Boris Szurek, Tuan Tu Tran, Hinda Doucoure, L.M. Jaimes Niño, Sébastien Cunnac |
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| Přispěvatelé: | UMR - Interactions Plantes Microorganismes Environnement (UMR IPME), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut de Recherche pour le Développement (IRD [France-Sud]), Universidad de los Andes [Bogota] (UNIANDES) |
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine [SDV.BIO]Life Sciences [q-bio]/Biotechnology Xanthomonas Bacterial leaf blight Clinical Biochemistry 01 natural sciences Genome 03 medical and health sciences Type III effector Xanthomonas oryzae TAL effector [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN] TALome lcsh:Science Gene Genetics biology urogenital system Effector food and beverages biology.organism_classification [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology [SDV.BV.PEP]Life Sciences [q-bio]/Vegetal Biology/Phytopathology and phytopharmacy Medical Laboratory Technology genomic DNA 030104 developmental biology Enrichment cloning Bacterial leaf streak Type III secretion lcsh:Q Restriction digest Rice 010606 plant biology & botany |
| Zdroj: | MethodsX MethodsX, 2018, 5, pp.1027-1032. ⟨10.1016/j.mex.2018.08.014⟩ MethodsX, Vol 5, Iss, Pp 1027-1032 (2018) |
| ISSN: | 2215-0161 |
| DOI: | 10.1016/j.mex.2018.08.014 |
| Popis: | 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. Method name: TAL fishing, Keywords: Enrichment cloning, Type III secretion, Type III effector, TAL effector, TALome, Xanthomonas, Bacterial leaf blight, Bacterial leaf streak, Rice |
| Databáze: | OpenAIRE |
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