NR AWYB
AU Zhou,H.; Hickford,J.G.; Fang,Q.
TI Technical note: determination of alleles of the ovine PRNP gene using PCR-single-strand conformational polymorphism analysis.
QU Journal of Animal Science 2005 Apr; 83(4): 745-9
PT journal article
AB Susceptibility to scrapie in sheep is linked to variation at codons 136, 154, and 171 in the host prion protein gene (PRNP). A number of techniques are available for detecting these polymorphisms, but none allow for a rapid and accurate determination of genotype. Here we describe PCR coupled with single-strand conformational polymorphism (SSCP) analysis, which allows for the accurate identification of ovine PRNP alleles. A gene region including codons 136 to 171 was amplified by PCR, and the amplimers were then denatured and subjected to electrophoresis in a nondenaturing polyacrylamide gel. Nine unique SSCP patterns, representing nine different alleles of the ovine PRNP gene, could be resolved. A new polymorphism (I/T) at codon 142 also was detected. The profiles produced by SSCP allowed for the accurate differentiation of PRNP alleles and could be employed to genotype PRNP in sheep.
MH *Alleles; Amino Acid Sequence; Animals; Base Sequence; DNA Primers/chemistry; Genetic Predisposition to Disease/genetics; Genotype; Molecular Sequence Data; Polymerase Chain Reaction/standards/veterinary; *Polymorphism, Single-Stranded Conformational; PrPsc Proteins/chemistry/*genetics; Scrapie/*genetics; Sequence Alignment/veterinary; Sequence Analysis, Protein/veterinary; Sheep
AD Cell Biology Group, Agriculture and Life Sciences Division, Lincoln University, Canterbury, New Zealand.
SP englisch
PO USA