:: Volume 3, Issue 2 (Summer 2013) ::
J Fasa Univ Med Sci 2013, 3(2): 124-129 Back to browse issues page
Cloning and Expression of Leptospira LipL32 Antigen as a Candidate for Rapid Diagnosis
Nooshin Sohrabi 1, Shohreh Azimi2
1- Payam-e-Noor University , nsohrabi75@yahoo.com
2- Islamic Azad Universit
Abstract:   (53512 Views)
Background and Objective: Leptospirosis as an important emerging infectious zoonotic disease caused by spirochetes of the genus Leptospira. Given the low sensitivity and long duration of its culture, the diagnosis of leptospirosis is mainly based on serological methods. The microscopic agglutination test (MAT) is considered as the reference method. Because of the complexity of the MAT, there is an urgent need for the development of new reliable and rapid screening tests for leptospirosis. Major leptospiral outer membrane proteins (OMPs), present only in pathologic strains, could be regarded as a good candidate for diagnostic studies. Here we report the cloning and expression of LipL32, as a prominent immunogenic protein, in a prokaryotic system.
Materials and Methods: After the amplification of LipL32 gene, it was cloned into the pQE30 vector. The insertion of LipL32 gene into the vector was screened and confirmed with restriction analysis and sequencing. The recombinant plasmid was transformed into E. coli M15 strain, and the expressed protein was identified by SDS-PAGE and western blotting. This recombinant protein with 6× His-tagged sequence was purified using Ni-NTA affinity column chromatography.
Results: The results revealed that the selected gene was successfully cloned in pQE30 vector and recombinant protein (rLipL32) of approximately ~32 kDa was produced, purified and confirmed by western blotting.
Conclusion: This recombinant protein could be potentially used for the development of serodiagnosis tests for the diagnosis of leptospirosis in humans and animals.
Keywords: Leptospirosis, Recombinant Protein, Cloning, LipL32, Protein expression
Full-Text [PDF 2073 kb]   (3035 Downloads)    
Type of Study: Research | Subject: Cellular-Molecular Biology
Received: 2013/02/3 | Accepted: 2013/10/6 | Published: 2013/10/6

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Volume 3, Issue 2 (Summer 2013) Back to browse issues page