Tran T. T., Doucoure H., Hutin M., Nino L. M. J., Szurek Boris, Cunnac Sébastien, Koebnik Ralf. (2018). Efficient enrichment cloning of TAL effector genes from Xanthomonas. Methodsx, 5, p. 1027-1032. ISSN 2215-0161.
Titre du document
Efficient enrichment cloning of TAL effector genes from Xanthomonas
Tran T. T., Doucoure H., Hutin M., Nino L. M. J., Szurek Boris, Cunnac Sébastien, Koebnik Ralf
Source
Methodsx, 2018,
5, p. 1027-1032 ISSN 2215-0161
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.
Plan de classement
Sciences fondamentales / Techniques d'analyse et de recherche [020]
;
Sciences du monde végétal [076]
