Cloning, sequencing, and polymorphisms of the Wistar-Imamichi rat growth hormone gene using PCR-SSCP

Abstract

We successfully cloned the Wistar-Imamichi (WI) rat growth hormone gene (GenBank accession: GQ890681), which contained 5 exons and 4 introns. Using the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) technique, a novel missense substitution single nucleotide polymorphism was identified and tested for Hardy-Weinberg equilibrium with the χ(2) test. This gene fragment was investigated in 50 adult rats and 50 neonatal rats by PCR-SSCP. Results showed that only intron 4 had a polymorphic locus at the 97th nucleotide position from A (in the AA genotype) to C (in the BB genotype). Further statistical analysis showed that this locus was in Hardy-Weinberg equilibrium. These results suggested that this specific pathogen-free WI rat population, which was bred in the barrier system of the Research Center for Laboratory Animal Science of Inner Mongolia University, has high hereditary stability.