A simple polymerase chain reaction assay for genotyping the retinal degeneration mutation (Pdeb(rd1)) in FVB/N-derived transgenic mice.
FVB/N mice are one of the most common inbred strains for the generation of transgenic animals. This mouse strain is preferred for transgenesis because of its fertilized oocytes, which have unique pronuclei for microinjection, and its vigorous reproductive performance along with consistently large litter sizes. However, these inbred mice carry a retinal degeneration mutation caused by a proviral insertion into the Pdeb gene, encoding the beta subunit of cGMP phosphodiesterase. This mutation (Pdeb(rd1), formerly known as rd) results in postnatal rod photoreceptor degeneration and causes severe visual impairment, which may be relevant for behavioural and vision-related research. This deficit can be overcome by crossing these mice with other mouse strains carrying the wild-type allele at the Pdeb locus. We have devised a simple polymerase chain reaction (PCR)-based method for distinguishing between the mutant and the wild-type alleles, thus allowing the efficient monitoring of the Pdeb(rd1) mutation in FVB/N-derived transgenic mice prior to experimentation where visual deficit is expected to have an influence in the phenotype.
Giménez
E
E
Departamento de Biología Molecular y Celular, Centro Nacional de Biotecnologia, Madrid, Spain.
Montoliu
L
L
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000235:genetics*; D000483:Alleles; D000818:Animals; D003433:Crosses, Genetic; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005260:Female; D005838:Genotype; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012162:Retinal Degeneration / Q000235:genetics* / Q000662:veterinary*
D004247:DNA; D015106:3',5'-Cyclic-GMP Phosphodiesterases