NR AXQT
AU Lampo,E.; van Poucke,M.; Hugot,K.; Hayes,H.; van Zeveren,A.; Peelman,L.J.
TI Cloning and Transcription Profiling of Shadow of Prion Protein in Sheep
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.164
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: Although susceptibility for transmissible spongiform encephalopathies (TSEs) in sheep is known to be influenced by polymorphisms of the prion protein gene (PRNP), differences identified in the sequence of PRNP can not explain all the variations of this susceptibility. Therefore, there is growing interest in other genes that might be involved in the pathogenesis of TSE. One of these genes is shadow of prion protein (SPRN), a gene coding for a protein having remarkable similarities with the prion protein. Until now, SPRN has not been described in sheep, a highly relevant species in prion matters.
Aim: The aim of this research was to clone and characterize the SPRN gene in sheep.
Methods: A BAC mini-contig containing SPRN was built by chromosome walking and mapped by FISH. The SPRN sequence in sheep was then obtained by sequencing with primers based on the bovine SPRN sequence and the 3' end was determined by sequencing cDNA from cerebrum mRNA. In addition, the transcription profile of SPRN was determined by RT-PCR in 21 tissues.
Results: Comparative mapping revealed the presence of the genes ECHS1, PAOX, MTG1, SPRN, LOC619207, CYP2E1 and at least a part of SYCE1 in the BAC minicontig. The two most exterior BAC clones of this contig have been mapped by FISH on Oari22q24. By cloning and sequencing the SPRN gene, a fragment of 4,544 bp was obtained, covering the entire SPRN gene and 1206 bp of the promoter region (Acc. No. DQ870545). SPRN in sheep contains 2 exons (108 bp and 2495 bp) which are separated by an intron of 735 bp. The coding region, entirely located in exon 2, codes for a protein of 146 amino acids and shares 93% respectively 78% nucleic acid identity, and 95% respectively 76% amino acid identity with that of cow and man. The transcription profile of SPRN in sheep was determined by RT-PCR, showing high levels of SPRN mRNA in cerebrum and cerebellum, and low levels in testis, lymph node, jejunum, ileum, colon and rectum.
Conclusion: Annotation of a mini-contig containing SPRN suggests conserved linkage between Oari22q24 and Hsap10q26. The ovine SPRN sequence, described for the first time, shows a high level of sequence identity with the bovine, and to a lesser extent with the human SPRN sequence. In addition, transcription profiling in sheep reveals expression of SPRN mainly in brain tissue, as in cow, man, rat and mouse.
AD E. Lampo, M. van Poucke, A. van Zeveren, L.J. Peelman, Faculty of Veterinary Medicine, Ghent University, Belgium; K. Hugot, INRA, Laboratoire de Radiobiologie et d'Etude du Genome, France; H. Hayes, INRA, Unité de Génétique Biochimique et Cytogénétique, France
SP englisch
PO Schottland