Molecular cloning and sequence analysis of prion protein gene in Xiji donkey in China

Abstract

Prion diseases are a group of human and animal neurodegenerative disorders caused by the deposition of an abnormal isoform prion protein (PrP(Sc)) encoded by a single copy prion protein gene (PRNP). Prion disease has been reported in many herbivores but not in Equus and the species barrier might be playing a role in resistance of these species to the disease. Therefore, analysis of genotype of prion protein (PrP) in these species may help understand the transmission of the disease. Xiji donkey is a rare species of Equus not widely reared in Ningxia, China, for service, food and medicine, but its PRNP has not been studied. Based on the reported PrP sequence in GenBank we designed primers and amplified, cloned and sequenced the PRNP of Xiji donkey. The sequence analysis showed that the Xiji donkey PRNP was consisted of an open reading frame of 768 nucleotides encoding 256 amino acids. Amino acid residues unique to donkey as compared with some Equus animals, mink, cow, sheep, human, dog, sika deer, rabbit and hamster were identified. The results showed that the amino acid sequence of Xiji donkey PrP starts with the consensus sequence MVKSH, with almost identical amino acid sequence to the PrP of other Equus species in this study. Amino acid sequence analysis showed high identity within species and close relation to the PRNP of sika deer, sheep, dog, camel, cow, mink, rabbit and hamster with 83.1-99.7% identity. The results provided the PRNP data for an additional Equus species, which should be useful to the study of the prion disease pathogenesis, resistance and cross species transmission.

Keywords: BSE; CJD; CWD; Creutzfeldt–Jakob disease; FME; GPI; ORF; PCR; PRNP; PrP; PrP(C); PrP(Sc); Prion gene; SNPs; TME; TSE; TSEs; Xiji donkey; bovine spongiform encephalopathy; cellular prion protein; chronic wasting disease; feline spongiform encephalopathy; glycosyl-phosphatidyl inositol; mink spongiform encephalopathy; open reading frame; polyenzyme chain reaction; prion protein; prion protein gene; scrapie isoform of PrP; single-nucleotide polymorphisms; transmissible spongiform encephalopathies; vCJD; variant Creutzfeldt–Jakob diseases.