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Protein & Peptide Letters
ISSN (Print): 0929-8665
ISSN (Online): 1875-5305
Epub Full Text Article
DOI:      Price:  $95









Cloning and Purification of IpaC Antigen from Shigella flexneri: Proposal of a New Methodology

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Author(s): Cristiane Mobilon, Marcelo Augusto Szymanski de Toledo, Fernanda Laroza Paganelli, Clelton Aparecido dos Santos, Fernanda de Pace, Jacqueline Boldrin de Paiva, Eliana Guedes Stehling, Gerson Nakazato, Aline G. Balieiro, Flávia P.S. Airoldi, Francisco de A. M. Reis and Wanderley Dias da Silveira
Abstract:
Shigella flexneri is a Gram-negative bacillus that is responsible for a severe form of dysentery called Shigellosis, which mainly affects children and the elderly in both underdeveloped and developed countries. Pathogenic S. flexneri strains possess a large virulence plasmid that codes for effector proteins that are required for the entry and spread of the bacteria into colonocytes. Among these proteins is the translocator IpaC, which plays an important role in the invasion process; IpaC is implicated in pore formation in the host cell membrane and induces cytoskeletal rearrangements in macrophages and epithelial cells, thereby promoting bacterial entry. The ability of IpaC to insert into the plasma membrane is due to a large nonpolar region of the protein structure. This characteristic also renders difficulties in recovery and purification when the protein is expressed in E. coli. Several works have considered different methodologies for the improved production and purification of IpaC. Herein, we propose an alternative method that is based on changes in the induction temperature and extraction buffer to facilitate the accumulation of high yields of soluble proteins for their further processing and ultimate use in biotechnological approaches.
Keywords:
Shigella flexneri, IpaC production, purification, shigellosis, vaccines
Affiliation:
Institute of Biology, PC6109, Department of Genetics, Evolution and Bioagents, State University of Campinas, São Paulo, Brazil