ARS | Publication request: A rapid sequence-based method for comprehensive polymorphism identification within a 25.2-kb region of the bovine prion gene in BSE-affected cattle

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: October 3, 2006
Publication Date: January 2, 2007
Citation: Clawson, M.L., Heaton, M.P., Keele, J.W., Smith, T.P., Harhay, G.P., Laegreid, W.W. 2007. A rapid sequence-based method for comprehensive polymorphism identification within a 25.2-kb region of the bovine prion gene in BSE-affected cattle [abstract]. Plant and Animal Genome XV Conference. Poster No. P533.

Technical Abstract: Bovine spongiform encephalopathy (BSE) is a fatal neurological disorder characterized by abnormal deposits of a protease-resistant isoform of the prion protein. Typical and atypical BSEs have been identified in cattle and the relationships of prion gene (PRNP) variation with susceptibility to these BSEs may not be identical. Thus, a robust approach to genotyping PRNP polymorphisms is required. Recently, we sequenced a 25.2-kb genomic region containing PRNP in 192 diverse U.S. beef and dairy cattle and observed 388 total polymorphisms. The relatively high density of PRNP polymorphisms, coupled with multiple repetitive elements distributed throughout the gene present a challenge for accurate PRNP amplification and genotyping. In this report, we describe an improved set of oligonucleotide primers that will reliably amplify the 25.2-kb PRNP region on 24 overlapping amplicons and provide comprehensive sequence coverage of the amplicons. The primer set can be used to 1) genotype the 388 known polymorphisms and 2) identify previously unknown polymorphisms within the 25.2-kb region. This strategy can be applied to the DNA of typical or atypical BSE cases to quickly identify PRNP polymorphisms.