A rapid, polymerase chain reaction-based procedure for identifying mutant restricted ovulator chickens

Females of the restricted ovulator (RO) strain of White Leghorn chickens fail to lay eggs upon photostimulation and exhibit endogenous hyperlipidemia and atherosclerotic lesions. A mutation in the gene specifying the oocyte vitellogenesis receptor (OVR), a 95-kDa membrane protein that normally media...

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Bibliographic Details
Published in:Poultry science 1996-09, Vol.75 (9), p.1113-1117
Main Authors: Bujo, H. (University of Vienna, Vienna, Austria.), Elkin, R.G, Lindstedt, K.A, Nimpf, J, Bitgood, J.J, Schneider, W.J
Format: Article
Language:English
Subjects:
Animals
Avian Proteins
Base Sequence
Chickens - genetics
Chickens - physiology
CHIMIORECEPTEUR
CHROMOSOME
CROMOSOMAS
DNA - analysis
DNA - chemistry
DNA - genetics
DNA Primers - analysis
DNA Primers - chemistry
DNA Primers - genetics
EPREUVE SUR LA DESCENDANCE
Female
Genotype
Heterozygote
Male
Membrane Proteins - genetics
Membrane Proteins - physiology
Molecular Sequence Data
MUTACION
MUTATION
OOGENESIS
OVOGENESE
Ovulation - genetics
Ovulation - physiology
PCR
Pedigree
Phenotype
POLLO
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - veterinary
POULET
PRUEBAS DE DESCENDENCIA
QUIMIORECEPTORES
Receptors, Cell Surface
Receptors, LDL - genetics
Receptors, LDL - physiology
SECUENCIA NUCLEOTIDICA
SEQUENCE NUCLEOTIDIQUE
SEXE
SEXO
TECHNIQUE ANALYTIQUE
TECNICAS ANALITICAS
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Summary:Females of the restricted ovulator (RO) strain of White Leghorn chickens fail to lay eggs upon photostimulation and exhibit endogenous hyperlipidemia and atherosclerotic lesions. A mutation in the gene specifying the oocyte vitellogenesis receptor (OVR), a 95-kDa membrane protein that normally mediates the massive uptake of yolk precursors from the serum, is responsible for this abnormal phenotype. Because a single nucleotide substitution (G lead to C) is responsible for the defective OVR, a PCR-based procedure, described herein, was developed in order to provide a rapid and accurate method for identifying chickens possessing the mutant allele. Polymerase chain reaction-amplified fray meets of apparently identical size (- 400 bp) were obtained from genomic DNA using primer pairs specific for either the wild-type or mutant genes. Through cloning and sequencing of the PCR-amplified products, the fragment sizes were determined to be 413 bp each, which, included an intron sequence. Polymerase chain reaction-amplified genomic DNA from wild-type (ovr+/ovr+) males, heterozygous carrier (ovr+/ovr-) males, and wild-type (-/ovr+) females all yielded a 413 bp fragment when a primer pair specific for the wild-type gene was used. Because female chickens are heterogametic (ZW), no PCR product was observed in the case of the mutant (-/ovr-) females. When the primer pair specific for the mutant gene was employed, PCR-amplification of genomic DNA from both heterozygous carrier (ovr+/ovr-) males and mutant (-/ovr-) females, but not wildtype (ovr+/ovr+) males or (-/ovr+) females, also yielded a 413-bp fragment. Employment of the present rapid and accurate procedure would be expected to obviate the need for conventional progeny testing while reducing the time required to identify RO carrier males and mutant females from approximately 1 yr to several days
ISSN:0032-5791
1525-3171
DOI:10.3382/ps.0751113