Abstract

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

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 χ2 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. 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 χ2 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. v

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