Advanced search
Start date
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

Full text
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]
Total Authors: 7
[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
Total Affiliations: 3
Document type: Journal article
Source: Protein Expression and Purification; v. 146, p. 78-84, JUN 2018.
Web of Science Citations: 1

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)

FAPESP's process: 13/50724-5 - Protein Kinase Chemical Biology Center: supporting drug development through open-access research
Grantee:Paulo Arruda
Support Opportunities: Research Grants - Research Partnership for Technological Innovation - PITE
FAPESP's process: 12/07657-2 - PR-1 proteins and new receptor like kinases of cacao (Theobroma cacao): gene expression evaluation and search for ligands
Grantee:Jorge Maurício Costa Mondego
Support Opportunities: Regular Research Grants