A sequencing strategy for identifying variation throughout the prion gene of BSE-affected cattle.

BMC Research Notes
Michael L ClawsonWilliam W Laegreid

Abstract

Classical and atypical bovine spongiform encephalopathies (BSEs) are cattle prion diseases. Distinct bovine prion gene (PRNP) alleles have been associated with classical and atypical BSE susceptibility. However, the full extent of PRNP allele association with BSE susceptibility is not known. A systematic sequence-based genotyping method that detects variation throughout PRNP would be useful for: 1) detecting rare PRNP alleles that may be present in BSE-affected animals and 2) testing PRNP alleles for an association with either classical or atypical BSE susceptibility. We improved a Sanger-based sequencing strategy for detecting bovine PRNP variation through all exons, introns, and part of the promoter (25.2 kb). Our current method can detect 389 known and other potentially unknown PRNP polymorphisms that may be present in BSE-affected cattle. We determined PRNP genotypes for the first U.S. BSE case and her sire. Previously unknown PRNP polymorphisms were not detected in either animal and all PRNP genotypes support the sire-daughter relationship. The methodologies described here characterize variation throughout PRNP. Consequently, rare PRNP alleles that may be present in BSE-affected cattle can be detected.

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Citations

May 4, 2010·BMC Veterinary Research·Michael P HeatonWilliam W Laegreid
Sep 24, 2010·PloS One·Brenda M MurdochStephen S Moore

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Datasets Mentioned

BETA
EU130450
EU130451

Methods Mentioned

BETA
genotyping
PCR
PCRs

Software Mentioned

Polyphred
Phred
Phrap
Consed

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