Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori
Objective(s): Helicobacter pylori, a human specific gastric pathogen is a causative agent of chronic active gastritis. The vacuolating cytotoxin (VacA) is an effective virulence factor involved in gastric injury. The aim of this study was to construct a recombinant protein containing antigenic regio...
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| Language: | English |
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Mashhad University of Medical Sciences
2013-07-01
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| Series: | Iranian Journal of Basic Medical Sciences |
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| Online Access: | http://ijbms.mums.ac.ir/pdf_1118_73ee7ed2b4d28e2eff03394946d803a0.html |
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| author | Leila Hasanzadeh Ehsanollah Ghaznavi-Rad Safieh Soufian Vahideh Farjadi Hamid Abtahi |
| author_facet | Leila Hasanzadeh Ehsanollah Ghaznavi-Rad Safieh Soufian Vahideh Farjadi Hamid Abtahi |
| author_sort | Leila Hasanzadeh |
| collection | DOAJ |
| description | Objective(s): Helicobacter pylori, a human specific gastric pathogen is a causative agent of chronic active gastritis. The vacuolating cytotoxin (VacA) is an effective virulence factor involved in gastric injury. The aim of this study was to construct a recombinant protein containing antigenic region of VacA gene and determine its antigenicity. Materials and Methods: The antigenic region of VacA gene was detected by bioinformatics methods. The polymerase chain reaction method was used to amplify a highly antigenic region of VacA gene from chromosomal DNA of H. pylori. The eluted product was cloned into the prokaryotic expression vector pET32a. The target protein was expressed in the Escherichia coli BL21 (DE3) pLysS. The bacteria including pET32a-VacA plasmids were induced by IPTG. The antigenicity was finally studied by western blotting using sera of 15 H. pylori infected patients after purification. Results: Enzyme digestion analysis, PCR and DNA sequencing results showed that the target gene was inserted correctly into the recombinant vector. The expressed protein was purified successfully via affinity chromatography. Data indicated that antigenic region of VacA protein from Helicobacter pylori was recognized by all 15 patient’s sera. Conclusion : Our data showed that antigenic region of VacA protein can be expressed by in E. co.li. This protein was recognized by sera patients suffering from H. pylori infection. the recombinant protein has similar epitopes and close antigenic properties to the natural form of this antigen. Recombinant antigenic region of VacA protein also seems to be a promising antigen for protective and serologic diagnosis . |
| first_indexed | 2024-04-12T18:39:02Z |
| format | Article |
| id | doaj.art-e21d0c876fdc4cfd9e4a3ec00679fe4b |
| institution | Directory Open Access Journal |
| issn | 2008-3866 2008-3874 |
| language | English |
| last_indexed | 2024-04-12T18:39:02Z |
| publishDate | 2013-07-01 |
| publisher | Mashhad University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Basic Medical Sciences |
| spelling | doaj.art-e21d0c876fdc4cfd9e4a3ec00679fe4b2022-12-22T03:20:51ZengMashhad University of Medical SciencesIranian Journal of Basic Medical Sciences2008-38662008-38742013-07-011678358401118Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter PyloriLeila Hasanzadeh0Ehsanollah Ghaznavi-Rad1Safieh Soufian2Vahideh Farjadi3Hamid Abtahi4¹ Department of Biotechnology and Microbiology, School of Medicine, Arak University of Medical Sciences, Arak, IranDepartment of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran³Biology Department, Payame Noor University, Arak, IranDepartment of Microbiology, Islamic Azad University, Qom Branch, Qom, IranMolecular and Medicine Research Center, Department of Microbiology, School of Medicine, Arak University of Medical Sciences, Arak, IranObjective(s): Helicobacter pylori, a human specific gastric pathogen is a causative agent of chronic active gastritis. The vacuolating cytotoxin (VacA) is an effective virulence factor involved in gastric injury. The aim of this study was to construct a recombinant protein containing antigenic region of VacA gene and determine its antigenicity. Materials and Methods: The antigenic region of VacA gene was detected by bioinformatics methods. The polymerase chain reaction method was used to amplify a highly antigenic region of VacA gene from chromosomal DNA of H. pylori. The eluted product was cloned into the prokaryotic expression vector pET32a. The target protein was expressed in the Escherichia coli BL21 (DE3) pLysS. The bacteria including pET32a-VacA plasmids were induced by IPTG. The antigenicity was finally studied by western blotting using sera of 15 H. pylori infected patients after purification. Results: Enzyme digestion analysis, PCR and DNA sequencing results showed that the target gene was inserted correctly into the recombinant vector. The expressed protein was purified successfully via affinity chromatography. Data indicated that antigenic region of VacA protein from Helicobacter pylori was recognized by all 15 patient’s sera. Conclusion : Our data showed that antigenic region of VacA protein can be expressed by in E. co.li. This protein was recognized by sera patients suffering from H. pylori infection. the recombinant protein has similar epitopes and close antigenic properties to the natural form of this antigen. Recombinant antigenic region of VacA protein also seems to be a promising antigen for protective and serologic diagnosis .http://ijbms.mums.ac.ir/pdf_1118_73ee7ed2b4d28e2eff03394946d803a0.htmlAntigenic region Cloning Epitopes Helicobacter pylori VacA cytotoxin |
| spellingShingle | Leila Hasanzadeh Ehsanollah Ghaznavi-Rad Safieh Soufian Vahideh Farjadi Hamid Abtahi Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori Iranian Journal of Basic Medical Sciences Antigenic region Cloning Epitopes Helicobacter pylori VacA cytotoxin |
| title | Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori |
| title_full | Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori |
| title_fullStr | Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori |
| title_full_unstemmed | Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori |
| title_short | Expression and Antigenic Evaluation of VacA Antigenic Fragment of Helicobacter Pylori |
| title_sort | expression and antigenic evaluation of vaca antigenic fragment of helicobacter pylori |
| topic | Antigenic region Cloning Epitopes Helicobacter pylori VacA cytotoxin |
| url | http://ijbms.mums.ac.ir/pdf_1118_73ee7ed2b4d28e2eff03394946d803a0.html |
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