MOLECULAR CLONING AND EXPRESSION OF THE LEISHMANIA TROPICA KMP-11 GENE | ||||
| Journal of the Egyptian Society of Parasitology | ||||
| Article 4, Volume 44, Issue 2, August 2014, Page 321-328 PDF (275.79 K) | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/jesp.2014.90383 | ||||
 View on SCiNiTO | ||||
| Authors | ||||
| MOUAYAD MERIEE1; CHADI SOUKKARIEH1; ABDUL QADER A. ABBADY2 | ||||
| 1Department of Animal Biology, Faculty of Sciences, Damascus University, Syria. | ||||
| 2Department of Molecular Biology and Biotechnology, Atomic Energy Commission of Syria (AECS), Damascus, Syria. | ||||
| 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. | ||||
| Keywords | ||||
| Leishmania tropica; Molecular cloning; Expression | ||||
|
Statistics Article View: 139 PDF Download: 177 |
||||
