title: Molecular Cloning and Characterization of P4 Nuclease from Leishmania infantum.
abstract: Parasite of the genus Leishmania is reliant on the salvage pathway for recycling of ribonucleotides. A class I nuclease enzyme also known as P4 nuclease is involved in salvage of purines in cutaneous Leishmania species but the relevant enzymes have not been characterized in Leishmania infantum (L. infantum). The aim of this study was to clone and characterize the gene encoding class I nuclease in L. infantum. DNA extracted from L. infantum was used for amplification of P4 nuclease gene (Li-P4) by PCR. The product was cloned, sequenced, and expressed in E. coli for further characterization. Analysis of the sequence of Li-P4 revealed that the gene consists of an ORF of 951 bp. Sequence similarity analysis indicated that Li-P4 has a high homology to relevant enzymes of other kintoplastids with the highest homology (88%) to p1/s1 class I nuclease from L. donovani. Western blotting of antirecombinant Li-P4 with promastigote and amastigote stages of L. infantum showed that this nuclease is present in both stages of parasite with higher expression in amastigote stage. The highly conserved nature of this essential enzyme in Leishmania parasites suggests it as a promising drug target for leishmaniasis.
authors.affiliation: Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz 51656-65811, Iran.
authors.forename: Safar
authors.initials: S
authors.lastname: Farajnia
authors.forename: Leila
authors.initials: L
authors.lastname: Rahbarnia
authors.forename: Bahram
authors.initials: B
authors.lastname: Maleki Zanjani
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Alimohammadian
authors.forename: Shahin
authors.initials: S
authors.lastname: Abdoli Oskoee
authors.forename: Abbas
authors.initials: A
authors.lastname: Beh-Pajooh
authors.forename: Nazli
authors.initials: N
authors.lastname: Saeedi
authors.forename: Soheila
authors.initials: S
authors.lastname: Montazer Saheb