Mammalian orthologous PCR of the putative TM4 of the NRAMP1 gene and molecular characterisation of this region in dogs

Orthologous gene-specific PCR primers have been designed to amplify the putative transmembrane domain (TM) 4 of the Nramp1 gene in mammals. Sequencing of the dog PCR fragment showed to be Nramp1 gene and it was used as a probe for Southern blot analysis, confirming the specific amplification of this gene also in other species. In order to analyse the relationship between Nramp and Leishmaniosis in dogs, a PCR-RFLP protocol has been optimised to look for TM4 polymorphism. Results showed a conserved Gly169residue in all animals tested and only two intronic polymorphisms.