title: Molecular cloning and biologically active production of IpaD N-terminal region.
abstract: Shigella is known as pathogenic intestinal bacteria in high dispersion and pathogenic bacteria due to invasive plasmid antigen (Ipa). So far, a number of Ipa proteins have been studied to introduce a new candidate vaccine. Here, for the first time, we examined whether the N-terminal region of IpaD(72-162) could be a proper candidate for Shigella vaccine. Initially, the DNA sequence coding N-terminal region was isolated by PCR from Shigella dysenteriae type I and cloned into pET-28a expression vector. Then, the heterologous protein was expressed, optimized and purified by affinity Ni-NTA column. Western blot analysis using, His-tag and IpaD(72-162) polyclonal antibodies, confirmed the purity and specificity of the recombinant protein, respectively. Subsequently, the high immunogenicity of the antigen was shown by ELISA. The results of the sereny test in Guinea pigs showed that IpaD(72-162) provides a protective system against Shigella flexneri 5a and S. dysenteriae type I.
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Hesaraki
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Saadati
authors.forename: Hossein
authors.initials: H
authors.lastname: Honari
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Olad
authors.forename: Mohammad
authors.initials: M
authors.lastname: Heiat
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Malaei
authors.forename: Reza
authors.initials: R
authors.lastname: Ranjbar
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D003001:Cloning, Molecular; D004405:Dysentery, Bacillary / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*; D015870:Gene Expression; D006168:Guinea Pigs; D008297:Male; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D022281:Shigella Vaccines / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D012762:Shigella dysenteriae; D012763:Shigella flexneri
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D022281:Shigella Vaccines
keywords: IpaD; Protectivity; Sereny test; Shigella dysenteriae