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Cloning and purification of IpaC antigen from shigella flexneri: Proposal of a new methodology
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نویسنده
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mobilon c. ,de toledo m.a.s. ,paganelli f.l. ,dos santos c.a. ,de pace f. ,de paiva j.b. ,stehling e.g. ,nakazato g. ,balieiro a.g. ,da silva airoldi f.p. ,de assis machado reis f. ,da silveira w.d.
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منبع
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protein and peptide letters - 2013 - دوره : 20 - شماره : 2 - صفحه:133 -139
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چکیده
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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. © 2013 bentham science publishers.
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کلیدواژه
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IpaC production; Purification; Shigella flexneri; Shigellosis; Vaccines
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آدرس
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institute of biology,department of genetics,evolution and bioagents,state university of campinas,campinas, Brazil, center of molecular biology and genetics engineering,state university of campinas,campinas, Brazil, department of medical microbiology,university medical center utrecht, Netherlands, center of molecular biology and genetics engineering,state university of campinas,campinas, Brazil, department of internal medicine,state university of campinas,campinas, Brazil, institute of biology,department of genetics,evolution and bioagents,state university of campinas,campinas, Brazil, department of toxicological and bromatologic clinical analyses,university of são paulo,ribeirão preto, Brazil, department of microbiology,londrina state university,londrina, Brazil, department of organic chemistry,institute of chemistry,state university of campinas,campinas, Brazil, department of organic chemistry,institute of chemistry,state university of campinas,campinas, Brazil, department of organic chemistry,institute of chemistry,state university of campinas,campinas, Brazil, institute of biology,department of genetics,evolution and bioagents,state university of campinas,campinas, Brazil
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Authors
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