A robust, low- to medium-throughput prnpgenotyping system in sheep

Buitkamp, Johannes; Semmer, Jördis

doi:10.1186/1471-2334-4-30

Cited by 12 publications

(8 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PCR-SSCP and ARMS-PCR have been used to detect genotypes (Buitkamp and Semmer, 2004;Zhou et al, 2005). In a previous study, amplified products of the unique SSCP patterns were cloned into a T-vector and were then sequenced (Zhou et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

“…Genotyping results proved that our method is much more suitable in this study because of the superiority in analyzing different genotypes with similar banding patterns. We also improved the previous ARMS-PCR (Buitkamp et al, 2004) by developing a series of easier test procedures but not labeling primer with fluorochromes. Our improved ARMS-PCR detected mutations at codons 136, 154 and 171 such as 136T, 154L and 171K, which could not be detected by the previous method described by Buitkamp et al (2004).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Ovine prion protein genotype frequencies in northwestern China

Zhao¹,

Wü²,

Liu³

et al. 2012

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Anti-scrapie breeding programs have been initiated to screen for scrapie-resistant sheep based on ovine prion protein gene (PRNP) genotypes at codons 136, 154 and 171 in many countries, especially European Union member states. However, investigation of sheep PRNP genotypes is limited in China, despite the large number of sheep breeds. We analyzed 432 sheep of five different breeds from farms in northwestern China, using PCR-single-strand conformational polymorphism analysis (PCR-SSCP); the corresponding haplotypes of different PRNP alleles were cloned. PRNP allele genotyping was done by amplification refractory mutation system-PCR (ARMS-PCR), according to the haplotype clones of each PRNP allele. The validity of ARMS-PCR was checked by PCR-SSCP. Another 325 unknown PRNP genotypes of other sheep breeds were analyzed according to the established ARMS-PCR. Genotype frequencies of 757 sheep were analyzed with these two methods to evaluate susceptibility to scrapie in northwestern China. Relevant mutations were also detected at other sites. Both methods were effective for ovine PRNP allele genotyping, and the results of the analysis completely coincided. Scrapie-resistant genotypes were found to be uncommon, indicating a high risk for ovine scrapie in northwest China. In addition to codons 136, 154 and 171, we found numerous new mutations; nearly half of them were previously unreported. These sheep populations have a high degree of polymorphism at the PRNP locus.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Ovine prion protein genotype frequencies in northwestern China

Zhao¹,

Wü²,

Liu³

et al. 2012

Genet. Mol. Res.

View full text Add to dashboard Cite

show abstract

“…The first attempts at the molecular discrimination of BSE and scrapie in sheep were made in the late 1990s and early 2000s (Bosser et al, 2000;Buitkamp and Semmer, 2004). Thanks to monoclonal antibodies obtained over the years for different epitopes of the prion protein, differences were detected in the pathological material from natural cases of ovine scrapie and experimental infections of sheep with BSE (Hill et al, 1998;Kuczius et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

Frequency of genotypes in the PrP prion protein gene locus in the polish sheep population

Rejduch

Knapik

Piestrzyńska-Kajtoch

et al. 2009

Acta Veterinaria Hungarica

View full text Add to dashboard Cite

Scrapie is an invariably fatal transmissible neurodegenerative disease of sheep, goats and moufflons, characterised by ataxia, lower body weight and changes in behaviour. The aim of this study was to determine the genotype distribution at codons 136, 154 and 171 of the PrP locus in sheep bred in Poland. The genotypes of 801 sheep representing 10 different breeds and crossbreds were analysed using real-time PCR allele discrimination method. The combination of point mutations of the three codons (136, 154 and 171) created twelve different genotypes in the PrP locus classified into five categories of risk. The highest frequency for the ARR/ARR genotype was characteristic of the Pogórze and Berrichon du Cher breeds (72.22% and 63.25%, respectively). In the Romanov breed (1.52%) and in crossbred sheep (2.61%) the VRQ/VRQ genotype was observed.Key words: Sheep, PrP gene, molecular methods, frequency of genotypes, frequency of alleles Scrapie is an invariably fatal transmissible neurodegenerative disease of sheep, goats and moufflons, characterised by ataxia, lower body weight and changes in behaviour. Other transmissible spongiform encephalopathies (TSEs) are bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in deer, and Creutzfeldt-Jacob disease (including vCJD and nvCJD) in humans (Goldmann et al., 2005). The lesions appear mainly in the nervous system in the form of vacuoles triggered by the conversion of the cellular prion pro-

show abstract

“…The DDEF1 SNP chosen using the Fisher exact test (DDEF1g.279401A>G, NCBI_ss181800428) was genotyped in 987 animals according to an adaptation of the ARMS methodology (Buitkamp and Semmer, 2004) and analyzed in an ABI 3100 Avant automatic DNA sequencer (Applied Biosystems).…”

Section: Snp's Genotyping In the Populationmentioning

confidence: 99%

Prospecting candidate SNPs for backfat in Canchim beef cattle

Veneroni

Meirelles²,

Grossi³

et al. 2010

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Canchim is a composite cattle breed developed in Brazil for beef production. One of the breeding objectives is to increase fat deposition. QTLs for fat thickness and/or marbling have been reported on BTA4 and BTA14. The IGFBP3 and DDEF1 genes, mapped to BTA4 and BTA14, respectively, affect adipogenesis. We looked for SNPs in the IGFBP3 and DDEF1 genes that could be associated with backfat thickness in Canchim beef cattle. For SNP identification, sires with the highest accuracy were ranked according to expected breeding value for fat thickness; the 12 extremes (six sires with the highest and six with the lowest expected breeding value for the trait) were chosen. Six regions of the IGFBP3 and 14 regions of the DDEF1 were sequenced using the Sanger method. Nine SNPs were identified in IGFBP3 and 76 in the DDEF1. After an initial analysis, two SNPs were selected to be genotyped for the whole population; these were DDEF1g.279401A>G and IGFBP3c.4394T>C(Trp>Arg). We found a significant effect (P ≤ 0.05) of allele substitution on backfat thickness; however, the IGFBP3 SNP did not significantly affect this trait.

show abstract

A robust, low- to medium-throughput prnpgenotyping system in sheep

Cited by 12 publications

References 16 publications

Ovine prion protein genotype frequencies in northwestern China

Ovine prion protein genotype frequencies in northwestern China

Frequency of genotypes in the PrP prion protein gene locus in the polish sheep population

Prospecting candidate SNPs for backfat in Canchim beef cattle

Contact Info

Product

Resources

About