Volume 3, Issue 1 (8-2016)                   J Jiroft Univ Med Sci 2016, 3(1): 82-91 | Back to browse issues page

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Kheiri-Dastenaei S, Doosti A. Cloning and evaluation of expression of the Helicobacter pylori ureB gene by using pcDNA3.1(+) expression vector. J Jiroft Univ Med Sci. 2016; 3 (1) :82-91
URL: http://journal.jmu.ac.ir/article-1-90-en.html
1- Department of Biology, Faculty of Basic Science,Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.
2- Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran. , abbasdoosti@yahoo.com
Abstract:   (3398 Views)

Introduction: Helicobacter pylori causes the gastritis, peptic ulcer, gastric cancer and lymphoma. There is no effective vaccine against this bacterium. Urease which is coded by ureB is one of the important stimulating agents for host immune system. The aim of this study was cloning and expression study of the ureB gene in order to create a gene vaccine against H. pylori.

Methods: In this experimental study, ureB gene was reproduced from Helicobacter pylori genome using PCR method. The PCR products were T/A cloned into pTZ vector and then digested with XhoI and XbaI enzymes and inserted to the pcDNA3.1(+) vector. pcDNA3.1(+)-ureB final construct was transformed in CHO cells using electroporation method. ureB gene expression was shown on a SDS-PAGE gel.

Results: The results show that the pTZ-ureB recombinant vector was generated. The restriction digest and sequencing confirmed the correctness of the cloning of ureB into pcDNA 3.1(+) vector. The electroporation results and study of gene expression in animal cells showed the desirable protein band on SDS-PAGE gel.

Conclusion: According to the results, the pcDNA3.1(+)-ureB expression vector can express the ureB gene product in eukaryotic cells. So the pcDNA3.1(+)-ureB final construct has a high potential as a DNA vaccine candidate for the evaluation of the immunogenicity in animal model.

Keywords: Helicobacter pylori, Recombinant vaccine, Gene cloning, ureB gene.

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Type of Study: Research | Subject: Medical Sciences / Medical Genetics
Received: 2016/11/15 | Accepted: 2016/12/8 | Published: 2016/12/20

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