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J. Anim. Sci. 2005. 83:745-749
© 2005 American Society of Animal Science

ANIMAL GENETICS

Technical Note: Determination of alleles of the ovine PRNP gene using PCR–single-strand conformational polymorphism analysis1

H. Zhou, J. G. H. Hickford2 and Q. Fang

Cell Biology Group, Agriculture and Life Sciences Division, Lincoln University, Canterbury, New Zealand

2 Correspondence: P. O. Box 84 (phone: +64-3-325 2811; fax: +64-3-325 3851; e-mail: hickford{at}lincoln.ac.nz).

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.

Key Words: Polymorphism • Prion Protein Gene (PRNP) • Sheep • Single-Strand Conformational Polymorphism




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