Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM

Sr35, a coiled-coil nucleotide-binding leucine-rich repeat receptor (CC-NLR) from the wheat species Triticum monococcum can directly recognize the pathogen avirulence factor AvrSr35 and confers immunity against wheat stem rust caused by Puccinia graminis f.sp. tritici race Ug99. Assembly of a stable...

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Autores principales: Liu, Meng-Xi, Zhao, Yan-Bo, Li, Ze-Kai, Chen, Tao-Tao, Ouyang, Songying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9637819/
https://www.ncbi.nlm.nih.gov/pubmed/36353364
http://dx.doi.org/10.1016/j.xpro.2022.101796
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author Liu, Meng-Xi
Zhao, Yan-Bo
Li, Ze-Kai
Chen, Tao-Tao
Ouyang, Songying
author_facet Liu, Meng-Xi
Zhao, Yan-Bo
Li, Ze-Kai
Chen, Tao-Tao
Ouyang, Songying
author_sort Liu, Meng-Xi
collection PubMed
description Sr35, a coiled-coil nucleotide-binding leucine-rich repeat receptor (CC-NLR) from the wheat species Triticum monococcum can directly recognize the pathogen avirulence factor AvrSr35 and confers immunity against wheat stem rust caused by Puccinia graminis f.sp. tritici race Ug99. Assembly of a stable Sr35 resistosome induced by AvrSr35 in vitro is usually limited by protein expression and low assembly efficiency. Here, we describe the expression and purification of AvrSr35 and Sr35, in vitro assembly of Sr35 resistosome for structure determination by cryo-EM. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2022).
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spelling pubmed-96378192022-11-08 Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM Liu, Meng-Xi Zhao, Yan-Bo Li, Ze-Kai Chen, Tao-Tao Ouyang, Songying STAR Protoc Protocol Sr35, a coiled-coil nucleotide-binding leucine-rich repeat receptor (CC-NLR) from the wheat species Triticum monococcum can directly recognize the pathogen avirulence factor AvrSr35 and confers immunity against wheat stem rust caused by Puccinia graminis f.sp. tritici race Ug99. Assembly of a stable Sr35 resistosome induced by AvrSr35 in vitro is usually limited by protein expression and low assembly efficiency. Here, we describe the expression and purification of AvrSr35 and Sr35, in vitro assembly of Sr35 resistosome for structure determination by cryo-EM. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2022). Elsevier 2022-10-31 /pmc/articles/PMC9637819/ /pubmed/36353364 http://dx.doi.org/10.1016/j.xpro.2022.101796 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Liu, Meng-Xi
Zhao, Yan-Bo
Li, Ze-Kai
Chen, Tao-Tao
Ouyang, Songying
Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM
title Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM
title_full Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM
title_fullStr Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM
title_full_unstemmed Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM
title_short Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM
title_sort assembly and purification of avrsr35-induced sr35 resistosome and determination of its structure by cryo-em
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9637819/
https://www.ncbi.nlm.nih.gov/pubmed/36353364
http://dx.doi.org/10.1016/j.xpro.2022.101796
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