Abstract
Assignment of the G3m(g) and (b) correlative amino acid residues was performed at the genomic level by direct sequencing of DNA from nine Caucasian individuals. Two oligonucleotide primers were used for subclass-specific enzymatic amplification of a DNA segment comprising a major portion of the second and third constant region domains (CH2 and CH3) fo the human IgG3 heavy chain gene. Comparison of the sequences of amplified DNA from individuals serologically typed as homozygous for G3m(b) or G3m(g) or as heterozygous, G3m(b,g), revealed differences in the codons for the amino acid residues 291, 296, and 384. Proline, phenylalanine, and serine at these positions corresponded to G3m(b), and leucine, tyrosine, and asparagine to G3m(g). Heterozygotic individuals, typed G3m(b,g), displayed both the G3m(b) and G3m(g) codons at these three positions. The polymorphism at each of these three codons could be identified either as the appearance, or the loss, of recognition sites for the two restriction endonucleases, Nsp BII and Rsa I. This allowed the development of a polymerase chain reaction (PCR)-based assay permitting the distinction of G3m b and G3m g alleles by analyzing the electrophoretical mobility of the DNA fragments generated by digestion of the PCR-products with Nsp BII and Rsa I.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Balbín, M., Grubb, A., Abrahamson, M., and Grubb, R. Determination of allotypes G1m(f) and (z) at the genomic level by subclass specific amplification of DNA and use of allele specific probes. Expl Clin Immunogenet 8: 88–95, 1991
Grey, H. M. and Abel, C. A. Subclass and allotype differences in human IgG immunoglobulins reflected by the amino acid sequence of Fc glycopeptides. Protides Biol Fluids 17: 229–231, 1970
Grubb, R. The genetic markers of human immunoglobulins. In A. Kleinzeller, G. F. Springer, and H. G. Whitman (eds.): Molecular Biology, Biochemistry and Biophysics, no. 9, Springer, Berlin Heidelberg New York, 1970
Grubb, R. Human immunoglobulin allotypes and Mendelian polymorphisms of the human immunoglobulin genes. In C. J. van Oss and M. H. V. van Regenmortel (eds.): Immunochemistry, Marcel Dekker, New York, in press
Grubb, R., Abrahamson, M., and Grubb, A. Assignment of allotypes G1m(a+) and G1m(a−) at the genomic level by polymerase chain reaction analysis. Expl Clin Immunogenet 7: 205–212, 1990
Grubb, R., Eberhardt, K., and Johnson, U. Alloimmunization to human immunoglobulin genetic markers is frequent in early rheumatoid artritis. Expl Clin Immunogenet 8: 219–226, 1991
Huck, S., Fort, P., Crawford, D. H., Lefranc, M.-P., and Lefranc, G. Sequence of a human immunoglobulin gamma 3 heavy chain constant region gene: comparison with other Cγ genes. Nucl Acids Res 14: 1779–1789, 1986
Ito, S., Yamazaki, K., Miyazaki, T.,and Matuumoto, H. An amino acid substitution determining G3m(g). J Biochem 99: 1799–1802, 1986
Ito, S., Suzuki, K., Miyazaki, T., and Matsumoto, H. Amino acid substitution determining G3m(g5). Expl Clin Immunogenet 7 : 91–100, 1990
Ito, S., Suzuki, K., Miyazaki, T., and Matsumoto, H. A key amino acid determining G3m(b) allotypic markers. Jpn J Human Genet 36: 179–187, 1991
Kabat, E. A., Wu, T. T., Reid-Miller, M., Perry, H. M., and Gottesman, K. S. Sequences of proteins of immunological interest, 4th edn, US Department of Health and Human Services, NIH, Bethesda, 1987
Matsumoto, H., Ito, S., Miyazaki, T., and Ohta, T. Structural studies of a human g3 myeloma protein (Jir) bearing the allotypic marker Gm(st). J Immunol 131: 1865–1870, 1983
Matsumoto, H. Characteristics of Mongoloid populations and immunogenetics of various diseases based on the genetic markers of human immunoglobulins. Expl Clin Immunogenet 6: 68–87, 1989
Nakao, Y. and Sasazuki, T. Gm and disease. In D. M. Weir (ed.): Handbook of Experimental Immunology, vol. 3: Genetics and Molecular Immunology, pp. 96.1–96.10, Blackwell Scientific Publications, Oxford and Edinburgh, 1986
Recht, B., Frangione, B., Franklin, E., and Van Loghem, E. Structural studies of a human γ3 myeloma protein (GOE) that binds Staph. Protein A. J Immunol 127: 917–923, 1981
Rosenbaum, V. and Riesner, D. Temperature gradient gel electrophoresis. Thermodynamic analysis of nucleic acids and proteins in purified form and in cellular extracts. Biophys Chem 26: 235–246, 1987
Schanfield, M. S. and van Loghem, E. In D. M. Weir (ed.) Handbook of Experimental Immunology, vol. 3: Genetics and Molecular Immunology, pp. 94.1–94.18, Blackwell Scientific Publications, Oxford and Edinburgh, 1986
Steinberg, A. G. and Cook, C. E. The distribution of the human immunoglobulin allotypes. Oxford Monographs on Medical Genetics, Oxford University Press, Oxford, 1981
Williams, R. C., Malone, C. C.,and Casali, P. Heteroclitic polyclonal and monoclonal anti-Gm(a) and aanti-Gm(g) human rheumatoid factors react with epitopes induced in Gm(a-), Gm(g-) by interaction with antigen or by nonspecific aggregation. J Immunol 149: 1817–1824, 1992
Author information
Authors and Affiliations
Additional information
Correspondence to: R. Grubb.
Rights and permissions
About this article
Cite this article
Balbín, M., Grubb, A., de Lange, G.G. et al. DNA sequences specific for Caucasian G3m(b) and (g) allotypes: allotyping at the genomic level. Immunogenetics 39, 187–193 (1994). https://doi.org/10.1007/BF00241259
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00241259