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. 2013 Nov;15(11):e8793.
doi: 10.5812/ircmj.8793. Epub 2013 Nov 5.

Cloning and Sequence Analysis of LipL32, a Surface-Exposed Lipoprotein of Pathogenic Leptospira Spp

Ebrahim Khodaverdi Darian  1 , Mohammad Mahdi Forghanifard  2 , Soheila Moradi Bidhendi  3 , Yung-Fu Chang  4 , Emad Yahaghi  5 , Majid Esmaelizad  6 , Maryam Khaleghizadeh  2 , Pejvak Khaki  3
Affiliations

Cloning and Sequence Analysis of LipL32, a Surface-Exposed Lipoprotein of Pathogenic Leptospira Spp

Ebrahim Khodaverdi Darian et al. Iran Red Crescent Med J. 2013 Nov.

Abstract

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.

Objectives: 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.

Materials and 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.

Keywords: Leptospira Interrogans; Leptospirosis; LipL32.

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Figures

Figure 1.
Figure 1.. PCR amplification of the 835 bp LipL32 gene of L.interrogans serovars: 100 bp DNA ladder, Positive control: serovar Pomona, Lane1: serovar Grippotyphosa, Lane 2: serovar Sejroe Hardjo, Lane3: serovar Canicola, Lane 4: saprophytic serovar Biflexa, and negative control.
Figure 2.
Figure 2.. Sequence Pair Distances of LipL32 Gene Sequences of Different Leptospiral Serovars
Figure 3.
Figure 3.. Phylogenetic Tree Analysis of LipL32 Gene
See this image and copyright information in PMC

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