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Cloning, expression and purification of Tex protein from Xanthomonas citri subsp. citri

Grant number: 11/23563-5
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): March 01, 2012
Effective date (End): December 31, 2012
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Jesus Aparecido Ferro
Grantee:Mariane Cristina Do Nascimento
Home Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

Brazil is the largest producer of orange and frozen concentrated orange juice in the world and São Paulo State is responsible for 80% of the orange and 98% of the juice produced in the country. One major problem of the citriculture worldwide is the diseases that attack the orchards and causes great economic losses. Citrus canker, caused by the Gram-negative bacteria Xanthomonas citri subsp. citri (Xac) is one of the major citrus disease and there is no effective control of the disease, with the only alternative being eradication.The genome sequencing of Xac opened the possibility of using functional genomics platforms to characterize genes involved in virulence and pathogenicity. The understanding of the mechanisms involved in Xac-citrus interaction is crucial to the development of strategies to control citrus canker.In a preliminary study, the disruption of ORF XAC2053 (tex gene) from Xac by transposon Tn5 mutagenesis resulted in a mutant with abolished canker symptoms in contrast to the wild type that causes citrus canker. The objective of the present study is to clone, express in E. coli and purify the Tex protein from Xac in a soluble form. The obtaining of the purified recombinant protein is the first step for further functional and structural studies in order to evaluate its potential as a target for the development of strategies to control the citrus canker disease.