Technical note: determination of alleles of the ovine PRNP gene using PCR-single-strand conformational polymorphism analysis.
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.
Zhou
H
H
Cell Biology Group, Agriculture and Life Sciences Division, Lincoln University, Canterbury, New Zealand.
Hickford
J G H
JG
Fang
Q
Q
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D020022:Genetic Predisposition to Disease / Q000235:genetics; D005838:Genotype; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000592:standards / Q000662:veterinary; D018807:Polymorphism, Single-Stranded Conformational; D018620:PrPSc Proteins / Q000737:chemistry / Q000235:genetics*; D012608:Scrapie / Q000235:genetics*; D016415:Sequence Alignment / Q000662:veterinary; D020539:Sequence Analysis, Protein / Q000662:veterinary; D012756:Sheep
D017931:DNA Primers; D018620:PrPSc Proteins