DNA sequences specific for Caucasian G3m(b) and (g) allotypes: allotyping at the genomic level |
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Authors: | Milagros Balbín Anders Grubb Gerda G de Lange Rune Grubb |
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Institution: | (1) Department of Clinical Chemistry, Lund University Hospital, S-22 185 Lund, Sweden;(2) Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, Holland;(3) Department of Medical Microbiology, University of Lund, Sölvegatan 23, S-22 362 Lund, Sweden;(4) Present address: Departamento de Biología Funcional, Area de Bioquímica, Universidad de Oviedo, 33006 Oviedo, Spain |
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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.
Correspondence to: R. Grubb. |
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