A C3H STRAIN-SPECIFIC ALLELE (ALPHA(VAL26)) OF THE MURINE ALPHA-GLOBIN GENE NEWLY DETECTED BY UT-PAGE AND RT-PCR-SSCP ANALYSIS

An extra band, distinct from the well-characterized globin chains (alp ha, beta(maj), beta(min), beta(s)), was detected in an adult erythrocy te sample of the C3H strain by urea triton polyacrylamide,eel electrop horesis PAGE) analysis. The extra band was recognized by an antibody a gainst the alpha-globin chain by Western blot analysis. Reverse transc ription, polymerase chain reaction and single strand conformation poly morphism (RT-PCR-SSCP) analysis and direct sequencing analysis of cDNA of the alpha-globin gene revealed a nucleotide substitution (GGA to G TA) corresponding to an amino acid substitution (Gly to Val) at codon 26 in the alpha-globin gene only in the erythrocyte sample of the C3H strain. Polypeptides generated by in vitro translation from the alpha- globin gene with the nucleotide substitution at codon 26 ((alpha(Val26 )) had the same mobility as that of the extra band of the C3H strain i n UT-PAGE. These results suggest that the substitution GGA (Gly) to GT A (Val) at codon 26 of the murine alpha-globin gene may directly affec t the mobility of alpha-globin in UT-PAGE and the base substitution ma y be a C3H strain-specific polymorphism.