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

Author
item Clawson, Michael - Mike
item Heaton, Michael - Mike
item Keele, John
item Smith, Timothy - Tim
item Harhay, Gregory
item Richt, Juergen
item Laegreid, William

Submitted to: BMC Research Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/23/2008
Publication Date: 6/23/2008
Citation: Clawson, M.L., Heaton, M.P., Keele, J.W., Smith, T.P., Harhay, G.P., Richt, J., Laegreid, W.W. 2008. A sequencing strategy for identifying variation throughout the prion gene of BSE-affected cattle. BMC Research Notes [journal online]. 1:32. Available: http://www.biomedcentral.com/1756-0500/1/32.

Interpretive Summary: Transmissible spongiform encephalopathies (TSEs) are fatal mammalian neurological disorders that are characterized by abnormal deposits of the prion protein. Distinct TSEs have been identified in humans, sheep and cattle. Classical bovine spongiform encephalopathy (BSE), identified during the mid-1980s, was the first TSE recognized in cattle and is the probable cause of the human TSE variant Creutzfeldt-Jakob Disease (vCJD). Recently, so-called "atypical" BSEs have been identified in North American and European cattle that have phenotypes different from classical BSE. Two atypical BSEs are known to afflict cattle. Variation in the bovine prion gene (PRNP) associates with susceptibility to classical BSE and may correlate with some atypical BSEs. Here, we present detailed methodologies for 1) efficiently extracting DNA from small portions of brainstem (obex) and 2) amplifying and sequencing the prion gene (promoter region through the 3’ untranslated region, 25.2 kb) with a primer set designed around known variation. Our method can be used to detect rare PRNP alleles that may be present in BSE-affected cattle and was applied to the U.S. classical BSE case of 2003 and her sire.

Technical Abstract: Cattle prion gene (PRNP) polymorphisms have been associated with bovine spongiform encephalopathy (BSE) susceptibility. We developed a method for sequencing bovine PRNP through all exons, introns and part of the promoter (25.2 kb) that accounts for known variation. The method can be used to detect rare PRNP alleles that may be present in BSE-affected cattle and was successfully applied to the first U.S. BSE case and her sire.