A C3H strain-specific allele (alpha va126) 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 (alpha, beta-maj, beta-min, beta-s), was detected in an adult erythrocyte sample of the C3H strain by urea triton polyacrylamide gel electrophoresis (UT-PAGE) analysis. The extra band was recognized by an antibody against the alpha-globin chain by Western blot analysis. Reverse transcription, polymerase chain reaction and single strand conformation polymorphism (RT-PCR-SSCP) analysis and direct sequencing analysis of cDNA of the alpha-globin gene revealed a nucleotide substitution (GGA to GTA) 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 in UT-PAGE. These results suggest that the substitution GGA (Gly) to GTA (Val) at codon 26 of the murine alpha-globin gene may directly affect the mobility of alpha-globin in UT-PAGE and the base substitution may be a C3H strain-specific polymorphism.
Sato
H
H
Tsukuba Life Science Center, Institute of Physical and Chemical Research, Ibaraki, Japan.
Nagayoshi
M
M
Ikawa
Y
Y
Kato
M V
MV
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000737:chemistry; D005914:Globins / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H / Q000235:genetics*; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D005914:Globins