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J D Works

Publications and source records attributed to J D Works.

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Molecular analysis and polymorphism of the DLA-DQB genes.

Partial-length cDNA clones and full-length genomic clones corresponding to a complete canine DQB class II gene were isolated. Southern analyses suggested the presence of two DQB genes--one of which appeared to be a pseudogene lacking exon 2 called DQB2. The other DQB gene, called DQB1, was isolated from a genomic phage clone and contained six exons. The DQB1 clone was restriction mapped, and exon 2 was sequenced from 70 dogs. Twenty alleles were found. Most of the amino acid substitutions occurred at putative positions in the peptide binding site. Inheritance of these sequences showed Mendelian segregation with one or two alleles per dog. Cluster analysis of the nucleotide and predicted amino acid sequences subdivided the canine DQB1 alleles into four major allelic groups. The number of nonsynonymous changes was higher than the number of synonymous changes in the putative antigen recognition sites suggestive of positive selection.

Amino Acid Sequence↗

DLA-DRB1 and DLA-DQB1 histocompatibility typing by PCR-SSCP and sequencing.

The dog has been an important model for solid organ and hematopoietic stem cell transplantation for over 30 years. Fundamental to the continuing usage of the model is the development of molecular-based histocompatibility typing of donors and recipients. Previous histocompatibility typing methods used in the dog have not been precise enough to identify dog leukocyte antigen (DLA)-matched unrelated dogs. This study was undertaken to begin the process of identifying DLA-matched unrelated dogs. In this study polymerase chain reaction-single-stranded conformational polymorphism is used to separate alleles thereby allowing sequenced-based typing of the two most polymorphic class II genes described to date in the dog DLA-DRB1 and DLA-DQB1.

Alleles↗

Molecular analysis of DLA-DRBB1 polymorphism.

The polymorphism of the canine major histocompatibility complex class II DRB gene, DRBB1, was analyzed. Exon 2 that encodes the polymorphic beta 1 domain was amplified by the polymerase chain reaction (PCR). The PCR product from 250 dogs was cloned and sequenced. Eighteen alleles were identified. Most of the variation in amino acid composition occurred at positions in the peptide binding site. Inheritance of these sequences showed Mendelian segregation with one or two alleles per dog. Cluster analysis of the nucleotide and predicted amino acid sequences subdivided the canine DRBB1 alleles into three major allelic groups. The number of nonsynonymous changes was higher than the number of synonymous changes in the putative antigen recognition sites indicative of positive selection. The data generated can serve as a basis for developing a typing assay for the canine DRBB1 gene.

Alleles↗