:: Volume 14, Issue 4 (Winter 2016) ::
jmj 2016, 14(4): 43-50 Back to browse issues page
Cloning of tagD gene from Helicobacter pylori in PFLAG-CMV-3 eukaryotic vector to generate a DNA vaccine
Maryam Safarpour, Zahra Kazemi, Elham Doosti, Abbas Doosti *
Islamic Azad University, Shahrekord Branch
Abstract:   (3972 Views)

Background: Helicobacter pylori is strongly associated with inflammation of the stomach and is also implicated in the development of gastric malignancy, peptic ulcers and gastric lymphomas in humans. Tpx (thiolperoxidases) is encoded by the tagD gene and plays a significant role in colonize of H. pylori to stomachs. This gene stimulates the immune system in the host cells. The aim of this study was to isolate and cloning of tagD gene in the eukaryotic expression vector PFLAG-CMV-3 as a DNA vaccine candidate.

Material & methods: In this research, tagD gene (537 bp) was amplified by PCR. The PCR products were cloned by using cloning kits of Thermo Fisher Company in pTZ vector. This gene was Subcloned in the eukaryotic expression vector (PFLAG-CMV-3 vector), then transferred into CHO cells by electroporation method and finally was expressed.

Results: The results indicate that the gene amplification and cloning of tagD gene, was successful, and the pTZ-tagD vector was formed. PFLAG-CMV-3 Vector construction was confirmed by digestion and gene sequencing. The 19 kDa band was observed by gene expression analysis on SDS-PAGE.

Conclusions: tagD gene in the PFLAG-CMV-3-tagD recombinant vector has an ability to produce specific protein in CHO cells. Therefore, this gene construct is useful to evaluate the immunogenicity as a DNA vaccine against of the Helicobacter pylori infection in human.

Keywords: Helicobacter pylori, tagD gene, cloning, electroporation
Full-Text [PDF 222 kb]   (1853 Downloads)    
Type of Study: Research | Subject: Microbshensi
Received: 2016/11/14 | Revised: 2017/07/11 | Accepted: 2017/03/13

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Volume 14, Issue 4 (Winter 2016) Back to browse issues page