MOLECULAR CLONING AND EXPRESSION OF THE LEISHMANIA TROPICA KMP-11 GENE

MERIEE, MOUAYAD; SOUKKARIEH, CHADI; ABBADY, ABDUL QADER A.

doi:10.21608/jesp.2014.90383

MOLECULAR CLONING AND EXPRESSION OF THE LEISHMANIA TROPICA KMP-11 GENE

Document Type : Original Article

Authors

¹ Department of Animal Biology, Faculty of Sciences, Damascus University, Syria.

² Department of Molecular Biology and Biotechnology, Atomic Energy Commission of Syria (AECS), Damascus, Syria.

10.21608/jesp.2014.90383

Abstract

Kinetoplastid membrane protein-11 (KMP-11) is a small protein of 11 kDa present in all kinetoplastid protozoa studded so far. This protein which is highly expressed in all stages of the Leishmania life cycle is considered a potential candidate for a leishmaniasis vaccine against many leishmania species. KMP-11 has been recently described in Leishmania tropica.
In the present study, the KMP-11 gene was extracted from L. tropica by PCR using two oligonucleotide primers designed to amplify the entire coding region of this gene. Then, the purified PCR products were successfully ligated into a high expression vector the pRSETGFP. This expression vector provides the opportunity to clone the desired insert as a fusion
protein with a GFP and a tag, polyhistidine region. The GFP use as a carrier to improve immune response and the polyhistidine tag facilitates detection of the expressed protein with anti-His antibodies and also purification of the protein using affinity purification. After wards KMP-11 coding region was sequenced and the recombinant protein was induced and
purified from Escherichia coli cultures. The results of the present study will increase our knowledge about molecular cloning and expression of the L. tropica KMP-11 gene, and this may be used as an effective target for controlling cutenous leishmaniasis.

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