Cloning, expression and purification of kinase domains of cacao PR-1 receptor-like kinases

Autor(es)

Tosarini, Thalita Ravazo ^[1] ; Ramos, Priscila Zonzini ^[2] ; Profeta, Gerson Souza ^[2] ; Baroni, Renata Moro ^[1] ; Massirer, Katlin B. ^{[2, 3]} ; Counago, Rafael M. ^{[2, 3]} ; Mondego, Jorge M. C. ^[1]

Afiliação do(s) autor(es)

^[1] Inst Agron Estado Sao Paulo, Campinas, SP - Brazil

^[2] Univ Campinas SGC UNICAMP, Struct Genom Consortium, Campinas, SP - Brazil

^[3] Univ Campinas CBMEG UNICAMP, Ctr Mol Biol & Genet Engn, Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Protein Expression and Purification; v. 146, p. 78-84, JUN 2018.
• Citações Web of Science: 1

Resumo

The PR-1 proteins (pathogenesis-related protein 1) are involved in plant defense mechanisms against various pathogens. The genome of cacao (Theobroma cacao) encodes 14 PR-1 proteins, named TcPR-1a to TcPR-1n. Two of them, TcPR-1f and TcPR-1g, have a C-terminal expansion with high similarity to protein kinase domains, suggesting a receptor-like kinase (RLK) protein architecture. Moreover, TcPR-1g is highly expressed during cacao response to Witches' Broom Disease, caused by the fungus Moniliopthora perniciosa. Here we describe a structural genomics approach to clone, express and purify the kinase domains of TcPR-1f and TcPR-1g. Escherichia coli BL21(DE3)-R3 cells were used for protein expression and co-expression of Lambda Protein Phosphatase was critical for successfully obtaining soluble recombinant protein. We expect that the ability to express and purify the kinase domains of TcPR-1f and TcPR-1g will further our understanding of the role these proteins play during cacao defense response. (AU)

• Processo FAPESP: 13/50724-5 - Centro de Biologia Química de Proteínas Quinases: alavancando desenvolvimento de fármacos através de pesquisa de acesso aberto
• Beneficiário: Paulo Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Parceria para Inovação Tecnológica - PITE
• Processo FAPESP: 12/07657-2 - Proteínas PR-1 e novos receptores do tipo quinase do cacaueiro (Theobroma cacao): avaliações de expressa gênica e busca por ligantes
• Beneficiário: Jorge Maurício Costa Mondego
• Modalidade de apoio: Auxílio à Pesquisa - Regular