Purification of effector-target protein complexes via transient expression in Nicotiana benthamiana.
Identifieur interne : 001739 ( Main/Corpus ); précédent : 001738; suivant : 001740Purification of effector-target protein complexes via transient expression in Nicotiana benthamiana.
Auteurs : Joe Win ; Sophien Kamoun ; Alexandra M E. JonesSource :
- Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2011.
English descriptors
- KwdEn :
- Agrobacterium tumefaciens (genetics), Chromatography, Liquid (methods), Gene Transfer Techniques (MeSH), Immunoprecipitation (methods), Phytophthora (genetics), Phytophthora (pathogenicity), Plant Immunity (MeSH), Tandem Mass Spectrometry (methods), Tobacco (genetics), Tobacco (metabolism), Tobacco (microbiology).
- MESH :
- genetics : Agrobacterium tumefaciens, Phytophthora, Tobacco.
- metabolism : Tobacco.
- methods : Chromatography, Liquid, Immunoprecipitation, Tandem Mass Spectrometry.
- microbiology : Tobacco.
- pathogenicity : Phytophthora.
- Gene Transfer Techniques, Plant Immunity.
Abstract
Effectors of plant pathogens play important roles in not only pathogenesis but also plant immunity. Plant pathogens use these effectors to manipulate host cells for colonization, and their activities likely influence the evolution of plant immune responses. Analyses of genome sequences revealed that oomycete pathogens, such as Phytophthora spp., possess hundreds of RXLR effectors that are thought to be delivered into the host cells and hence function inside the cells by interacting with the host protein complexes. This article describes a co-immunoprecipitation protocol aimed at identifying putative target complexes of the effectors by transiently overexpressing the tagged effectors in planta. The identification of the eluted protein complexes was achieved by LC-MS/MS mass spectrometry and peptide spectrum matching.
DOI: 10.1007/978-1-61737-998-7_15
PubMed: 21359809
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pubmed:21359809Le document en format XML
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Jones</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2011">2011</date><idno type="RBID">pubmed:21359809</idno><idno type="pmid">21359809</idno><idno type="doi">10.1007/978-1-61737-998-7_15</idno><idno type="wicri:Area/Main/Corpus">001739</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">001739</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Purification of effector-target protein complexes via transient expression in Nicotiana benthamiana.</title><author><name sortKey="Win, Joe" sort="Win, Joe" uniqKey="Win J" first="Joe" last="Win">Joe Win</name><affiliation><nlm:affiliation>The Sainsbury Laboratory, John Innes Centre, Norwich, UK.</nlm:affiliation></affiliation></author><author><name sortKey="Kamoun, Sophien" sort="Kamoun, Sophien" uniqKey="Kamoun S" first="Sophien" last="Kamoun">Sophien Kamoun</name></author><author><name sortKey="Jones, Alexandra M E" sort="Jones, Alexandra M E" uniqKey="Jones A" first="Alexandra M E" last="Jones">Alexandra M E. Jones</name></author></analytic><series><title level="j">Methods in molecular biology (Clifton, N.J.)</title><idno type="eISSN">1940-6029</idno><imprint><date when="2011" type="published">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Agrobacterium tumefaciens (genetics)</term><term>Chromatography, Liquid (methods)</term><term>Gene Transfer Techniques (MeSH)</term><term>Immunoprecipitation (methods)</term><term>Phytophthora (genetics)</term><term>Phytophthora (pathogenicity)</term><term>Plant Immunity (MeSH)</term><term>Tandem Mass Spectrometry (methods)</term><term>Tobacco (genetics)</term><term>Tobacco (metabolism)</term><term>Tobacco (microbiology)</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium tumefaciens</term><term>Phytophthora</term><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Chromatography, Liquid</term><term>Immunoprecipitation</term><term>Tandem Mass Spectrometry</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>Phytophthora</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Transfer Techniques</term><term>Plant Immunity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Effectors of plant pathogens play important roles in not only pathogenesis but also plant immunity. Plant pathogens use these effectors to manipulate host cells for colonization, and their activities likely influence the evolution of plant immune responses. Analyses of genome sequences revealed that oomycete pathogens, such as Phytophthora spp., possess hundreds of RXLR effectors that are thought to be delivered into the host cells and hence function inside the cells by interacting with the host protein complexes. This article describes a co-immunoprecipitation protocol aimed at identifying putative target complexes of the effectors by transiently overexpressing the tagged effectors in planta. 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Plant pathogens use these effectors to manipulate host cells for colonization, and their activities likely influence the evolution of plant immune responses. Analyses of genome sequences revealed that oomycete pathogens, such as Phytophthora spp., possess hundreds of RXLR effectors that are thought to be delivered into the host cells and hence function inside the cells by interacting with the host protein complexes. This article describes a co-immunoprecipitation protocol aimed at identifying putative target complexes of the effectors by transiently overexpressing the tagged effectors in planta. 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