Cloning and Sequence Analysis of LipL32, a Surface-Exposed Lipoprotein of Pathogenic Leptospira Spp.
BACKGROUND
Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira species. A major challenge of this disease is the application of basic research to improve diagnostic methods and related vaccine development. Outer membrane proteins of Leptospira are potential candidates that may be useful as diagnostic or immunogenic factors in treatment and analysis of the disease.


OBJECTIVE
To develop an effective subunit vaccine against prevalent pathogenic Leptospira species, we sequenced and analyzed the LipL32 gene from three different Leptospira interrogans (L.interrogans) vaccinal serovars in Iran.


METHODS
Following DNA extraction from these three serovars, the related LipL32 genes were amplified and cloned in the pTZ57R/T vector. Recombinant clones were confirmed by colony- PCR and DNA sequencing. The related sequences were subjected to homology analysis by comparing them to sequences in the Genbank database.


RESULTS
The LipL32 sequences were >94% homologous among the vaccinal and other pathogenic Leptospira serovars in GenBank. This result indicates the conservation of this gene within the pathogenic Leptospires.


CONCLUSIONS
The cloned gene in this study may provide a potentially suitable platform for development of a variety of applications such as serological diagnostic tests or recombinant vaccines against leptospirosis.
Young Researchers and Elite Club, Karaj Branch, Islamic Azad University, Karaj, IR Iran.
Ebrahim
E
Khodaverdi Darian
Department of Biology, Damghan Branch, Islamic Azad University, Damghan, IR Iran.
Mohammad Mahdi
MM
Forghanifard
Department of Microbiology, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
Soheila
S
Moradi Bidhendi
Department of Population, Medicine and Diagnostic sciences, College of Veterinary Medicine, Cornell University, Ithaca, USA.
Yung-Fu
YF
Chang
Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, IR Iran.
Emad
E
Yahaghi
Department of Biotechnology, Razi Vaccin and Serum Resaerch Institute, Karaj, IR Iran.
Majid
M
Esmaelizad
Department of Biology, Damghan Branch, Islamic Azad University, Damghan, IR Iran.
Maryam
M
Khaleghizadeh
Department of Microbiology, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
Pejvak
P
Khaki
Leptospira Interrogans; Leptospirosis; LipL32