caa a22 4500 39756855X CHVBK 20180308164834.0 cr unu---uuuuu 161202e199609 xx s 000 0 eng 10.3382/ps.0751113 doi (NATIONALLICENCE)oxford-10.3382/ps.0751113 A Rapid, Polymerase Chain Reaction-Based Procedure for Identifying Mutant Restricted Ovulator Chickens [Elektronische Daten] [H. BUJO, R. G. ELKIN, K. A. LINDSTEDT, J. NIMPF, J. J. BITGOOD, W. J. SCHNEIDERS] 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 → 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 fragments 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 wild-type (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. © 1996 Poultry Science Association, Inc. Molecular Biology nationallicence chicken nationallicence restricted ovulator nationallicence oocyte vitellogenesis receptor nationallicence polymerase chain reaction nationallicence single gene defect nationallicence BUJO H. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria, aut ELKIN R. G. Department of Animal Sciences, Purdue University, West Lafayette, Indiana 47907-1151, and aut LINDSTEDT K. A. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria, aut NIMPF J. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria, aut BITGOOD J. J. Department of Poultry Science, University of Wisconsin-Madison, Madison, Wisconsin 53706-1284 aut SCHNEIDERS W. J. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria, aut Poultry Science Oxford University Press 75/9(1996-09), 1113-1117 0032-5791 75:9<1113 1996 75 ps https://doi.org/10.3382/ps.0751113 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.3382/ps.0751113 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- BUJO H. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria aut NATIONALLICENCE 700 1- ELKIN R. G. Department of Animal Sciences, Purdue University, West Lafayette, Indiana 47907-1151, and aut NATIONALLICENCE 700 1- LINDSTEDT K. A. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria aut NATIONALLICENCE 700 1- NIMPF J. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria aut NATIONALLICENCE 700 1- BITGOOD J. J. Department of Poultry Science, University of Wisconsin-Madison, Madison, Wisconsin 53706-1284 aut NATIONALLICENCE 700 1- SCHNEIDERS W. J. Department of Molecular Genetics, Biocenter and University of Vienna, A-1030 Vienna, Austria aut NATIONALLICENCE 773 0- Poultry Science Oxford University Press 75/9(1996-09), 1113-1117 0032-5791 75:9<1113 1996 75 ps Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford