Cloning and Expression of Brucella Cyclic ?-1, 2 Glucan Transporter Gene (cgt)

AUTHORS

Mojgan Bandehpour 1 , Narges Abdali 2 , Farzaneh Sadeghi 3 , Kazem Parivar 2 , Zarrin Sharifnia 1 , Hossein Ryahi 3 , Ali Haghighi 4 , Bahram Kazemi 1 , *

1 Cellular and Molecular Biology Research Center, Shahid Beheshti University, M.C., Tehran, IR Iran

2 Islamic Azad University, Research and Science Campus, Tehran, IR Iran

3 Department of Biology, Shahid Beheshti University, M.C., Tehran, IR Iran

4 Department of Parasitology, Shahid Beheshti University, M.C., Tehran, IR Iran

How to Cite: Bandehpour M, Abdali N, Sadeghi F, Parivar K, Sharifnia Z, et al. Cloning and Expression of Brucella Cyclic ?-1, 2 Glucan Transporter Gene (cgt), Arch Clin Infect Dis. Online ahead of Print ; 4(1):3-7.

ARTICLE INFORMATION

Archives of Clinical Infectious Diseases: 4 (1); 3-7
Article Type: Research Article

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Abstract

Background: Brucellosis is an important cosmopolitan infection disease caused by organisms belonging to the genus Brucella. The cgt gene (cyclic ?-1, 2 glucan transporter gene) is a virulent factor in Brucella genus. The present study was conducted with the aim of cloning and expression of Brucella cgt gene.

Materials and methods: Brucella melitensis cgt gene was amplified from extracted chromosomal DNA by PCR, then

PCR product was cloned into pTZ57R and subcloned into pGEMEX-1 expression vector, then expressed in JM109 E.coli strain. Recombinant protein was confirmed by western blot analysis using patient's serum.

Results: The PCR product was cloned in pTZ57R plasmid via T/A cloning method. Recombinant plasmid was digested by BamHI and SacI restriction enzymes, the released band was purified and subcloned into pGEMEX-1 expression vector. Then, sample cells were lysed using lyses buffer and sonicated, then electrophoresed on SDS-PAGE. Protein bands were transferred on nitrocellulose membrane and reacted by patient's serum and detected by HRP conjugated anti

human antibody.

Conclusion: We cloned and expressed Brucella abortus cyclic -1, 2-glucan transporter gene (cgt) which is an important agent in brucellosis. Using cgt gene mutant may be an effective way for inhibiting or decreasing the pathogenicity of bacteria

Keywords

Brucella, Cyclic beta 1-2 glucan transporter, Recombinant protein.

© 0, Archives of Clinical Infectious Diseases. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

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