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Pakistan Journal of Biological Sciences
  Year: 2010 | Volume: 13 | Issue: 16 | Page No.: 775-784
DOI: 10.3923/pjbs.2010.775.784
 
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Molecular Cloning, Characterization and Overexpression of a Novel Cyclin from Leishmania mexicana

Nahla O.M. Ali, Muntaser E. Ibrahim, Karen M. Grant and Jeremy C. Mottram

Abstract:
We are reporting here, the cloning and characterization of the first cyclin from Leishmania mexicana. We have identified a cyclin-like motif from the L. major genome sequencing project. A cyclin homologue was cloned and sequenced from L. mexicana genome and it showed 96.1% amino acid identity with the putative L. major cyclin. It has also sequence identity to mitotic cyclins from other organisms. Southern analysis showed that it is present as a single copy gene. CYCa has been over-expressed in E. coli as a histidine fusion and western blot has confirmed the immunoreactive property of the recombinant cyclin, which then used to reconstitute active recombinant L. mexicana CRK3. No phosphorylation of histone HI was detected by both wild type and mutated CRK3 on the activation assays suggesting that phosphorylation status and cyclin binding are important for reconstituting protein kinase activity. The results confirm that we have isolated a cyclin molecule from L. mexicana (LmCYCa) which may play an important role in the regulation of the parasite cell cycle.
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How to cite this article:

Nahla O.M. Ali, Muntaser E. Ibrahim, Karen M. Grant and Jeremy C. Mottram, 2010. Molecular Cloning, Characterization and Overexpression of a Novel Cyclin from Leishmania mexicana. Pakistan Journal of Biological Sciences, 13: 775-784.

DOI: 10.3923/pjbs.2010.775.784

URL: https://scialert.net/abstract/?doi=pjbs.2010.775.784

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