Abstract
The objective of this study was to determine whether more than one DQB gene is expressed in three BoLA haplotypes that have a duplicated DQ region. Leukocyte mRNA from three animals genotyped at the BoLA-A, DQB, and DRB3 loci was used as template for reverse transcription-polymerase chain reaction (RT-PCR), cloning, and DNA sequencing. Five DQB alleles were identified. All cDNA clones were 564 base pairs (bp) in length, including 507 bp of nonprimer-derived sequence that contained the coding sequence for the full length of exon 2 and 74 amino acids of exon 3. Three alleles were assigned to the DQB1 locus and two were assigned to DQB2 on the basis of sequence comparisons with previously reported alleles. The expression of DQB1 and DQB2 in individual animals was examined by RT-PCR followed by double digestion of the 564 bp PCR products with Eco O109 I and Dra III in order to discriminate the DQB alleles. Evidence that DQB1 and DQB2 are both transcribed was obtained for three different BoLA haplotypes with DQB duplications, DQB10, DQB11C, and DQB12. These results suggest that the duplication or deletion event that gave rise to the DQB1 and DQB2 genes is a relatively recent event in the evolution of the cattle MHC.
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Ammer, H., Schwaiger, F.-W., Kammerbauer, C., Gomolka, M., Arriens, A., Lazary, S., and Epplen, J. T. Exonic polymorphism vs intronic simple repeat hypervariability in MHC-DRB genes. Immunogenetics 35: 332–340, 1992.
Andersson, L., Böhme, J., Peterson, P. A., and Rask, L. Genomic hybridization of bouvine class II major histocompatibility genes. 2. Polymorphism of DR genes and linkage disequilibrium in the DQ-DR region. Anim Genet 17: 295–304, 1986
Andersson, L., Lundén, A., Sigurdardóttir, S., Davies, C. J., and Rask, L. Linkage relationships in the bovine MHC region. High recombination frequency between class II subregions. Immunogenetics 27: 273–280, 1988
Andersson, L., and Rask, L. Characterization of the MHC class II region in cattle: the number of DQ genes varies between haplotypes. Immunogenetics 27: 110–120, 1988
Bernoco, D., Lewin, H. A., Andersson, L., Arriens, M. A., Cwik, S., Davies, C. J., Hines, H. C., Leibold, W., Lie, Ø., Meggiolaro, D., Østergaard, H., Spooner, R. L., Stewart-Haynes, J. A., Teale, A. J., Templeton, J. W., and Zanotti, M. Joint report of the Fourth International Bovine Lymphocyte Antigen (BoLA) Workshop, East Lansing, Michigan, USA, August 25, 1990. Anim Genet 22: 477–496, 1991
Bissumbhar, B., Joosten, I., Davis, W. C., and Hensen, E. J. Isoelectric focusing of bovine MHC DQ allelic variants. Anim Genet 23: 39, 1992
Burke, M. G., Stone, R. T., and Muggli-Cockett, N. E. Nucleotide sequences and Northern analysis of a bovine major histocompatibility class II DRβ-like cDNA. Anim Genet 22: 343–352, 1991
Glišin, V., Crkvenjakov, R., and Byus, C. Ribonucleic acid isolation by cesium chloride centrifugation. Biochemistry 13: 2633–2637, 1974
Groenen, M. A. M., van der Poel, J. J., Dijkhof, R. J. M., and Giphart, M. J. The nucleotide sequence of bovine MHC class II DQB and DRB genes. Immunogenetics 31: 37–44, 1990
Joosten, I., Sanders, M. F., van der Poel, A., Williams, J. I., Hepkema, B. G., and Hensen, E. J. Biochemically defined polymorphism of bovine MHC class II antigens. Immunogenetics 29: 213–216, 1989
Maniatis, T., Fritsch, E. F., and Sambrook, J. Molecular Cloning: A Laboratory Manua, pp. 239–326, Cold Spring Harbor Laboratory, Cold Spring Harbor, 1982
Muggli-Cockett, N. E. and Stone, R. R. Identification of genetic variation in the bovine major histocompatibility complex DRβ-like genes using sequenced bovine genomic probes. Anim Genet 19: 213–225, 1988
Muggli-Cockett, N. E. and Stone, R. T. Partial nucleotide sequence of a bovine major histocompatibility class II DRβ-like gene. Anim Genet 20: 361–369, 1989
Romano, M. J., Stewart, J. A., and Lewin, H. A. Phenotypic characterization of bovine lymphoblastoid cell lines. Vet Immunol Immunopathol 23: 293–307, 1989
Sanger, F., Nicklen, S., and Coulson, A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467, 1977
Shewey, L. M., Beaty, J. S., Andersen, L. C., and Nepom, G. T. Differential expression of related HLA class II DQβ genes caused by nucleotide variation in transcriptional regulatory elements. J Immunol 148: 1265–1273, 1992
Sigurdardóttir, S., Borsch, C., Gustafsson, K., and Andersson, L. Gene duplications and sequence polymorphism of bovine class II DQB genes. Immunogenetics 35: 205–213, 1992a
Sigurdardóttir, S., Borsch, C., Gustafsson, K., and Andersson, L. Exon encoding the antigen-binding site of MHC class II β-chains is divided into two subregions with different evolutionary histories. J Immunol 128: 968–973, 1992b
Sorrentino, R., Potolicchio, I., Ferrara, G. B., and Tosi, R. A new approach to HLA-DP1 typing combining DNA heteroduplex analysis with allele-specific amplification and enzyme restriction. Immunogenetics 36: 248–254, 1992
Spies, T., Sorrentino, R., Boss, J. M., Okada, K., and Strominger, J. L. Structural oragnization of the DR subregion of the human major histocompatibility complex. Proc Natl Acad Sci USA 82: 5165–5169, 1985
Teutsch, M. R., Stewart-Haynes, J. A., Beever, J. E., Xu, A., Schook, L. B., and Lewin, H. A. Haplotypes of the BoLA-A, Bf, CYP21 and DQB loci in Angus cattle. Animal Biotechnology 1: 185–199, 1990
Ullrich, A., Shine, J., Chirgwin, J., Pictet, R., Tischer, E., Rutter, W. J., and Goodman, H. M. Rat insulin genes: construction of plasmids containing the coding sequences. Science 196: 1313–1319, 1977
Van Eijk, M. J. T., Stewart-Haynes, J. A., and Lewin, H. A. Extensive polymorphism of the BoLA-DRB3 gene distinguished by PCR-RFLP. Anim Genet 23: 483–496, 1992
Van Eijk, M. J. T., Russ, I., and Lewin, H. A. Order of bovine DRB3, DYA and PRL determined by sperm typing. Mammalian Genome 4: 113–118, 1993
Watkins, D. I., Shadduck, J. A., Rudd, C. E., Stone, M. E., Lewin, H. A., and Letvin, N. L. Isoelectric focusing of bovine major histocompatibility complex class II molecules. Eur J Immunol 19: 567–570, 1989
Xu, A., Clark, T. J., Teutsch, M. R., Schook, L. B., and Lewin, H. A. Sequencing and genetic analysis of a bovine DQB cDNA clone. Anim Genet 22: 381–398, 1991
Xu, A., McKenna, K., and Lewin, H. A. Sequencing and genetic analysis of a bovine DQα cDNA clone. Immunogenetics 37: 231–234, 1993
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Xu, A., Park, C. & Lewin, H.A. Both DQB genes are expressed in BoLA haplotypes carrying a duplicated DQ region. Immunogenetics 39, 316–321 (1994). https://doi.org/10.1007/BF00189227
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DOI: https://doi.org/10.1007/BF00189227