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


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.


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


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

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