Schwaiger F W, Buitkamp J, Weyers E, Epplen J T
Molekulare Humangenetik, MA, Ruhr-Universität, Bochum, Germany.
Mol Ecol. 1993 Feb;2(1):55-9. doi: 10.1111/j.1365-294x.1993.tb00099.x.
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.
本文描述了一种针对牛、羊和山羊等偶蹄目动物高度多态的MHC-DRB基因的高效寡核苷酸分型方法。通过聚合酶链反应，扩增MHC-DRB的第二外显子以及包含混合简单重复序列的部分相邻内含子。使用这种引物组合，我们能够从所有被研究的10种牛科动物和长颈鹿中扩增出MHC-DRB外显子2和相邻内含子。因此，新的偶蹄目物种的DRB基因也能够很容易地被研究。使用针对有蹄类动物DRB基因多态性的寡核苷酸探针，可以区分至少一个单碱基不同的序列。发现外显子多态性与等位基因长度和重复结构模式相关。因此，针对不同简单重复和多态性位置的寡核苷酸探针也可用于跨物种分型。序列长度与外显子多态性的严格相关性允许预先选择用于杂交的特定寡核苷酸。因此，已经能够从这三个物种中的每一个中区分出20多个等位基因。
