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DNA sequences specific for Caucasian G3m(b) and (g) allotypes : allotyping at the genomic level

Balbín, Milagros ; Grubb, Anders LU orcid ; de Lange, Gerda G. and Grubb, Rune (1994) In Immunogenetics 39. p.187-193
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... (More)

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 G3mb and G3mg alleles by analyzing the electrophoretical mobility of the DNA fragments generated by digestion of the PCR-products with Nsp BII and Rsa I.

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author
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publishing date
type
Contribution to journal
publication status
published
in
Immunogenetics
volume
39
pages
187 - 193
publisher
Springer
external identifiers
  • pmid:8276465
  • scopus:0028084872
ISSN
0093-7711
DOI
10.1007/BF00241259
language
English
LU publication?
no
id
861fa749-909d-4c15-98ab-eb717a6134ee
date added to LUP
2021-10-28 13:14:35
date last changed
2024-01-12 02:58:40
@article{861fa749-909d-4c15-98ab-eb717a6134ee,
  abstract     = {{<p>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 (C<sub>H</sub>2 and C<sub>H</sub>3) 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<sup>b</sup> and G3m<sup>g</sup> alleles by analyzing the electrophoretical mobility of the DNA fragments generated by digestion of the PCR-products with Nsp BII and Rsa I.</p>}},
  author       = {{Balbín, Milagros and Grubb, Anders and de Lange, Gerda G. and Grubb, Rune}},
  issn         = {{0093-7711}},
  language     = {{eng}},
  pages        = {{187--193}},
  publisher    = {{Springer}},
  series       = {{Immunogenetics}},
  title        = {{DNA sequences specific for Caucasian G3m(b) and (g) allotypes : allotyping at the genomic level}},
  url          = {{http://dx.doi.org/10.1007/BF00241259}},
  doi          = {{10.1007/BF00241259}},
  volume       = {{39}},
  year         = {{1994}},
}