Application of Temperature-Gradient Gel Electrophoresis for Detection of Prion Protein Gene Polymorphisms in Polish Świniarka Sheep

Abstract

Abstract. This study presents preliminary data on the polymorphism in the prion protein gene of Ś winiarka sheep using temperature gradient gel electrophoresis (TGGE). Available data indicate that sensitivity to scrapie is associated with polymorphisms in three codons of prion protein gene: 136,154, and 171. The TGGE method was used to detect point mutations in these codons responsible for sensitivity or resistance to scrapie. This study revealed presence of an allele encoding valine (V) in codon 136, which is associated with high sensitivity to scrapie and occurred in the form of heterozygous allele together with alanine (AV). The highest variability was observed in codon 171, with presence of arginine (R) and glutamine (Q) in the homozygous (RR or QQ) as well as the heterozygous form (RQ). The results of examination of fifty sheep DNA samples with mutations in codons 136, 154, and 171 demonstrated that TGGE can be used as a simple and rapid method to detect mutations in the PrP gene of sheep. Several samples can be run at the same time, making TGGE ideal for the screening of large numbers of samples.

Prion Protein Coding Gene (PRNP) Variability in Sheep from Turkey and Iran

“…These include descriptions of Austrian (Sipos et al 2002), British (Arnold et al 2002;Baylis et al 2004), French (Palhière et al 2002), German (Drögemüller et al 2001(Drögemüller et al , 2004, Greek (Billinis et al 2004), Irish (O'Doherty et al 2000, Icelandic (Thorgeirsdottir et al 1999), Italian (Blasi et al 2003;Vaccari et al 2001), Norwegian (Tranulis et al 1999), Polish (Jasik and Reichert 2006), Portuguese (Gama et al 2006;Orge et al 2003), and Spanish (Acin et al 2004;Legarra et al 2004) breeds. The numerically predominant allele in all of the corresponding breeds is the tentative ancestral form of the PRNP gene with the ARQ allele.…”

Capillary electromigration based techniques in diagnostics of prion protein caused diseases

“…Recently, TGGE was used to find that sensitivity to scrapie is associated with polymorphisms in three codons of prion protein gene: 136, 154, and 171. The TGGE method was used to detect point mutations in these codons responsible for sensitivity or resistance to scrapie . In another recent study, the authors utilized recombinant human PrP as a probe in combination with 2DE and MALDI‐TOF MS for identification of heterogeneous nuclear ribonucleoprotein A2/B1 and aldolase C as novel interaction partners for PrP .…”

A decade of using small-to-medium throughput allele discrimination assay to determine prion protein gene (Prnp) genotypes in sheep in Slovenia

“…7 Various methods are available to determine the Prnp genotype, including restriction-fragment length polymorphisms, 8 sequencing, 12 dot-blot hybridization, 13 denaturing gradient gel electrophoresis analysis, 1 TaqMan assay, 3,8,[15][16][17] and temperature gradient gel electrophoresis. 14 We describe herein the results of our small-to-medium throughput allele discrimination assay as used in our laboratory beginning in 2006 to determine Prnp genotypes in sheep in Slovenia in the genotyping program.…”