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

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

Volume: 20 Issue: 2

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 Gambaro Balieiro, Flavia Pereira da Silva Airoldi, Francisco de Assis Machado Reis and Wanderley Dias da Silveira

Affiliation:

Keywords: Shigella flexneri, IpaC production, purification, shigellosis, vaccines

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.

Cite this article as:

Mobilon Cristiane, Augusto Szymanski de Toledo Marcelo, Laroza Paganelli Fernanda, Aparecido dos Santos Clelton, de Pace Fernanda, Boldrin de Paiva Jacqueline, Guedes Stehling Eliana, Nakazato Gerson, Gambaro Balieiro Aline, Pereira da Silva Airoldi Flavia, de Assis Machado Reis Francisco and Dias da Silveira Wanderley, Cloning and Purification of IpaC Antigen from Shigella flexneri: Proposal of a New Methodology, Protein & Peptide Letters 2013; 20 (2) . https://dx.doi.org/10.2174/0929866511320020003