A Rapid Screening Method by the Polymerase Chain Reaction-Single Strand Conformation Polymorphism Using Fluorescent Labeled Primer on Automatic Sequencer for Porcine Ryanodine Receptor Gene Mutation

抄録

A rapid screening method to detect the ryanodine receptor gene mutation (C1843 to T) for the porcine stress syndrome was developed by polymerase chain reaction-single strand conformation polymorphism using fluorescent labled primer on automatic sequencer. The 83bp of ryanodine receptor DNA fragment including mutated position was amplified from the porcine hair root using fluorescence labeled primer and electrophoresed on the 6% Hydrolink Long Ranger gel at 4°C. In this condition, the normal and mutant alleles were clearly separated. More than 35 samples were analyzed within 2 h and the gel was used several times daily. The 241 samples from 4 Japanese farms were analyzed by this method. The homozygous mutation was not found in all samples. But the high frequency (39.7% and 37.5%) of the heterozygous allele was observed in the two of the farm whilst the others were low (9.6% and 14.3%) . This method is suitable for analyzing a large number of samples in a short time to compare with halothane test and other genetic methods.