Typing of artiodactyl MHC-DRB genes with the help of intronic simple repeated DNA sequences.
An efficient oligonucleotide typing method for the highly polymorphic MHC-DRB genes is described for artiodactyls like cattle, sheep and goat. By means of the polymerase chain reaction, the second exon of MHC-DRB is amplified as well as part of the adjacent intron containing a mixed simple repeat sequence. Using this primer combination we were able to amplify the MHC-DRB exons 2 and adjacent introns from all of the investigated 10 species of the family of Bovidae and giraffes. Therefore, the DRB genes of novel artiodactyl species can also be readily studied. Oligonucleotide probes specific for the polymorphisms of ungulate DRB genes are used with which sequences differing in at least one single base can be distinguished. Exonic polymorphism was found to be correlated with the allele lengths and the patterns of the repeat structures. Hence oligonucleotide probes specific for different simple repeats and polymorphic positions serve also for typing across species barriers. The strict correlation of sequence length and exonic polymorphism permits a preselection of specific oligonucleotides for hybridization. Thus more than 20 alleles can already be differentiated from each of the three species.
Schwaiger
F W
FW
Molekulare Humangenetik, MA, Ruhr-Universität, Bochum, Germany.
Buitkamp
J
J
Weyers
E
E
Epplen
J T
JT
D000483:Alleles; D000818:Animals; D001188:Artiodactyla / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D004247:DNA / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes; D017753:Ecosystem; D005091:Exons; D006041:Goats; D007438:Introns; D008285:Major Histocompatibility Complex; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D012756:Sheep; D013045:Species Specificity
D017931:DNA Primers; D015342:DNA Probes; D004247:DNA