A protein expression system for tandem affinity purification in Xanthomonas citri subsp. citri
Identifieur interne : 000351 ( Main/Exploration ); précédent : 000350; suivant : 000352A protein expression system for tandem affinity purification in Xanthomonas citri subsp. citri
Auteurs : Giordanni C. Dantas [Brésil] ; Paula M. M. Martins [Brésil] ; Daniela A. B. Martins [Brésil] ; Eleni Gomes [Brésil] ; Henrique Ferreira [Brésil]Source :
- Brazilian Journal of Microbiology [ 1517-8382 ] ; 2016.
Abstract
Citrus canker, caused by the Gram-negative bacterium
Url:
DOI: 10.1016/j.bjm.2016.01.026
PubMed: 26991273
PubMed Central: 4874617
Affiliations:
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Martins</name><affiliation wicri:level="2"><nlm:aff id="aff0005">Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Martins, Daniela A B" sort="Martins, Daniela A B" uniqKey="Martins D" first="Daniela A. B." last="Martins">Daniela A. B. Martins</name><affiliation wicri:level="2"><nlm:aff id="aff0010">Depto. de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, Araraquara, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, Araraquara, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Gomes, Eleni" sort="Gomes, Eleni" uniqKey="Gomes E" first="Eleni" last="Gomes">Eleni Gomes</name><affiliation wicri:level="2"><nlm:aff id="aff0015">Depto. de Biologia, Universidade Estadual Paulista, São Jose do Rio Preto, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Biologia, Universidade Estadual Paulista, São Jose do Rio Preto, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Ferreira, Henrique" sort="Ferreira, Henrique" uniqKey="Ferreira H" first="Henrique" last="Ferreira">Henrique Ferreira</name><affiliation wicri:level="2"><nlm:aff id="aff0005">Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26991273</idno><idno type="pmc">4874617</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4874617</idno><idno type="RBID">PMC:4874617</idno><idno type="doi">10.1016/j.bjm.2016.01.026</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000E65</idno><idno type="wicri:Area/Pmc/Curation">000E64</idno><idno type="wicri:Area/Pmc/Checkpoint">000277</idno><idno type="wicri:Area/Ncbi/Merge">001F69</idno><idno type="wicri:Area/Ncbi/Curation">001F69</idno><idno type="wicri:Area/Ncbi/Checkpoint">001F69</idno><idno type="wicri:doubleKey">1517-8382:2016:Dantas G:a:protein:expression</idno><idno type="wicri:Area/Main/Merge">000351</idno><idno type="wicri:Area/Main/Curation">000351</idno><idno type="wicri:Area/Main/Exploration">000351</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A protein expression system for tandem affinity purification in <italic>Xanthomonas citri</italic> subsp. citri</title><author><name sortKey="Dantas, Giordanni C" sort="Dantas, Giordanni C" uniqKey="Dantas G" first="Giordanni C." last="Dantas">Giordanni C. Dantas</name><affiliation wicri:level="2"><nlm:aff id="aff0005">Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Martins, Paula M M" sort="Martins, Paula M M" uniqKey="Martins P" first="Paula M. M." last="Martins">Paula M. M. Martins</name><affiliation wicri:level="2"><nlm:aff id="aff0005">Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Martins, Daniela A B" sort="Martins, Daniela A B" uniqKey="Martins D" first="Daniela A. B." last="Martins">Daniela A. B. Martins</name><affiliation wicri:level="2"><nlm:aff id="aff0010">Depto. de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, Araraquara, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, Araraquara, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Gomes, Eleni" sort="Gomes, Eleni" uniqKey="Gomes E" first="Eleni" last="Gomes">Eleni Gomes</name><affiliation wicri:level="2"><nlm:aff id="aff0015">Depto. de Biologia, Universidade Estadual Paulista, São Jose do Rio Preto, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Biologia, Universidade Estadual Paulista, São Jose do Rio Preto, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Ferreira, Henrique" sort="Ferreira, Henrique" uniqKey="Ferreira H" first="Henrique" last="Ferreira">Henrique Ferreira</name><affiliation wicri:level="2"><nlm:aff id="aff0005">Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP, Brazil</nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Depto. de Bioquimica e Microbiologia, Instituto de Biociências, Universidade Estadual Paulista, Rio Claro, SP</wicri:regionArea><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author></analytic><series><title level="j">Brazilian Journal of Microbiology</title><idno type="ISSN">1517-8382</idno><idno type="eISSN">1678-4405</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Citrus canker, caused by the Gram-negative bacterium <italic>Xanthomonas citri</italic> subsp. citri (Xac), is one of the most devastating diseases to affect citrus crops. There is no treatment for citrus canker; effective control against the spread of Xac is usually achieved by the elimination of affected plants along with that of asymptomatic neighbors. An in depth understanding of the pathogen is the keystone for understanding of the disease; to this effect we are committed to the development of strategies to ease the study of Xac. Genome sequencing and annotation of Xac revealed that ∼37% of the genome is composed of hypothetical ORFs. To start a systematic characterization of novel factors encoded by Xac, we constructed integrative-vectors for protein expression specific to this bacterium. The vectors allow for the production of TAP-tagged proteins in Xac under the regulation of the xylose promoter. In this study, we show that a TAP-expression vector, integrated into the <italic>amy</italic> locus of Xac, does not compromise its virulence. Furthermore, our results also demonstrate that the polypeptide TAP can be overproduced in Xac and purified from the soluble phase of cell extracts. Our results substantiate the use of our vectors for protein expression in Xac thus contributing a novel tool for the characterization of proteins and protein complexes generated by this bacterium <italic>in vivo</italic>.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Da Silva, A C" uniqKey="Da Silva A">A.C. da Silva</name></author><author><name sortKey="Ferro, J A" uniqKey="Ferro J">J.A. Ferro</name></author><author><name sortKey="Reinach, F C" uniqKey="Reinach F">F.C. Reinach</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alegria, M C" uniqKey="Alegria M">M.C. Alegria</name></author><author><name sortKey="Docena, C" uniqKey="Docena C">C. Docena</name></author><author><name sortKey="Khater, L" uniqKey="Khater L">L. 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Butland</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Brésil</li></country><region><li>État de São Paulo</li></region></list><tree><country name="Brésil"><region name="État de São Paulo"><name sortKey="Dantas, Giordanni C" sort="Dantas, Giordanni C" uniqKey="Dantas G" first="Giordanni C." last="Dantas">Giordanni C. Dantas</name></region><name sortKey="Ferreira, Henrique" sort="Ferreira, Henrique" uniqKey="Ferreira H" first="Henrique" last="Ferreira">Henrique Ferreira</name><name sortKey="Gomes, Eleni" sort="Gomes, Eleni" uniqKey="Gomes E" first="Eleni" last="Gomes">Eleni Gomes</name><name sortKey="Martins, Daniela A B" sort="Martins, Daniela A B" uniqKey="Martins D" first="Daniela A. B." last="Martins">Daniela A. B. Martins</name><name sortKey="Martins, Paula M M" sort="Martins, Paula M M" uniqKey="Martins P" first="Paula M. M." last="Martins">Paula M. M. 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