NR AWPP
AU van Poucke,M.; Vandesompele,J.; De Ketelaere,A.; Goossens,K.; van Zeveren,A.; Peelman,L.J.
TI Prnp polymorphisms in sheep with differential Prnp mRNA expression in blood
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions GEN-29
PT Konferenz-Poster
AB As there is substantial evidence that substrate PrPc is required in specific tissues and cells for PrPsc accumulation to occur and because of the probable haematogenous spread of PrPsc throughout the lymphoreticular system during TSE pathogenesis in sheep, PRNP mRNA expression levels in sheep blood was investigated in 234 sheep. PRNP mRNA expression could be detected in only 11.5% of the samples, using a real-time quantitative PCR assay based on SYBR Green I detection, according to standard operating procedure guidelines. To correct for possible experimental variation, proper normalization of the cDNA input of all samples was achieved using 3 stably expressed reference genes (viz. RPL13A, RPS18 and UBC), chosen from a panel of 9 tested candidate reference genes using the geNorm algorithm. Our gene expression results show true biological variation of PRNP mRNA in sheep blood cells. To look for possible polymorphisms related to the observed variable expression, sequencing of 25,000 bp of the PRNP gene was carried out for 20 pairs of strongly related sheep with differential PrPc expression, constituting 7 different races (Ardense Voskop, Bleu du Maine, Hampshire Down, Rouge de l'Ouest, Suffolk, Texel and Vlaams Kuddeschaap) and the 5 predominant alleles (ARR, ARH, AHQ, ARQ and VRQ). In total, 283 polymorphisms were identified, of which 231 were not previously described. All polymorphisms are the subject of a genetic association study currently carried out. Some of these PRNP polymorphisms may not only elucidate the mechanisms controlling PrPc expression at the level of PRNP gene transcription and translation, but may also have an impact on TSE resistance.
AD M. Van Poucke, K. Goossens, A. Van Zeveren, L.J. Peelman: Department of Nutrition, Genetics and Ethology, Ghent University, Merelbeke, Belgium; J. Vandesompele: Centre for Medical Genetics, Ghent University Hospital, Ghent, Belgium; A. De Ketelaere: Department of Physiology, Biochemistry and Biometry, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium E-mail: Mario.VanPoucke@UGent.be
SP englisch
PO Italien