production of molecular clones of toxoplasma gondii-derived heat shock protein 70 kDa

Toxoplasmosis is a common and widespread infection in humans and many other species of warmblooded animals. Toxoplasma gondii-derived heat shock protein 70 [Hsp70] may play an important role in the virulence of Toxoplasma gondii [T. gondii]. In the present study, T. gondii Hsp70 was amplified by polymerase chain reaction [PCR] from the DNA of the T. gondiitachyzoite RH strain through the use of specific primers with Xho1 and Xba1 restriction sites. The purified DNA fragment of the T. gondii Hsp70 gene was subcloned into the Xho1 and Xba1-digested eukaryotic expression vector, pcDNA3, and subsequently transformed into TOP10 chemically competent cells. A 2004 base pair [bp] band of PCR product was observed on the 0.8% agarose gel. The cDNA was inserted into the pTZ57R/T vector and then subcloned successfully into the pcDNA3 eukaryotic expression plasmid vector. The sequence of this amplified gene showed up to 100% homology with the target gene according to the Genbank database [Accession no. U82281]

Detection and identification of Leishmania parasites sandflies using kDNA, rDNa and CPB loci

Molecular epidemiology of Visceral Leishmaniasis [VL] is currently used widely for different objectives such as vector incrimination studies. In this study three different loci including kinetoplast DNA [kDNA], ribosomal DNA [rDNA], cystein protease B [CPB[ of Leishmania parasite genome were used for detection and identification of natural infection of sand flies of Germi district of Ardebil province, the most important VL or Kala-azar foci in Iran. The results showed that the three loci of kDNA, rDNA and CPBs are respectively more appropriate for leptomonad infection/initial screening, identification of the L.donovani complex, and discrimination of the species complex. It was also verified that both members of the complex, L.donovani and L.infantum, are present in the study area and are transmitted to the hosts by Phlebotomus perfiliewi transcaucasicus sandflies. This is the first report on natural infection of sand flies to L.donovani in the country and since the ecology and biology of L.donovani differs extensively from L.infantum, it is necessary to perform further studies to highlight the role of L.donovani in epidemiology of VL in the region and country