Cloning and Expression of Helicobacter Pylori UreB122(a Segment of the B-subunit of Urease Gene)

Shahin Najar Peerayeh; Moein Farshchian; Majid Sadeghizadeh; Javad Atoofi

Archives of Clinical Infectious Diseases

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Cloning and Expression of Helicobacter Pylori UreB122(a Segment of the B-subunit of Urease Gene)

Author(s):

Shahin Najar Peerayeh^1,*,

Moein Farshchian¹,

Majid Sadeghizadeh²,

Javad Atoofi¹

1Department of Microbiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran

2Department of Genetic, School of Basic Sciences, Tarbiat Modares University, Tehran, IR Iran

Archives of Clinical Infectious Diseases:Vol. 6, issue 4; 161-4

Article type:Research Article

How to Cite:Shahin Najar PeerayehMoein FarshchianMajid SadeghizadehJavad AtoofiCloning and Expression of Helicobacter Pylori UreB122(a Segment of the B-subunit of Urease Gene).Arch Clin Infect Dis.6(4):161-4.

Abstract

Introduction:

Helicobacter pylori is associated with the chronic gastritis, peptic ulcer, gastric adenocarcinoma, and gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Moreover, antibiotic therapies do not protect against potential re-infection while have risk for development of drug resistance. Therefore, vaccine-mediated protection against H. pylori became an attractive clinical interest. H. pylori urease plays an important role in survival and pathogenesis of the infection. In this study the UreB122 (aa143-264) gene cloned in pET expression vector and the recombinant protein (rUreB122) was over expressed in Escherichia coli (E. coli).

Patients and Methods:

Genomic DNA of the standard H. pylori strain 26695 was isolated as the template and UreB122 gene was amplified by PCR. Prokaryote expression vector pET32a was inserted with UreB122 gene (pET32a-UreB122). The recombinant plasmid was used to transform competent E. coli DH5?, and positive clones were selected. Then the recombinant plasmid was used to transform E. coli BL21DE3 for expression of recombinant protein UreB122.The expression of recombinant protein was induced by isopropythio-?-D-galctoside (IPTG) at different concentration

Results:

In comparison with the reported corresponding sequences, the nucleotide sequence homology of UreB122 gene was 99.9%. UreB122 fusion protein was able to react with the anti His-Tag antibody .

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