Sauermann U, Arents A, Hunsmann G
German Primate Centre, Department of Virology and Immunology, Göttingen, Germany.
Tissue Antigens. 1996 Apr;47(4):319-28. doi: 10.1111/j.1399-0039.1996.tb02560.x.
Up to now 19 allelic sequences of the rhesus monkey DQB1 locus have been published. Referring to these sequences, we have developed a typing protocol for Mamu-DQB1 alleles which was verified by additional cloning, sequence analysis and segregation studies. The protocol is based on the amplification of the second exon with only one specific primer pair followed by the digestion of the PCR products with up to 10 different restriction endonucleases. The alleles can be identified in homozygous and heterozygous combinations since most amplified second exon sequences give unique hand patterns after digestion with at least one of the selected restriction endonucleases. By the use of this protocol we analyzed DNA-samples from 182 rhesus monkeys. Among these samples two novel Mamu-DQB1 alleles were detected, subsequently cloned and their nucleic sequence determined. Since we typed four complete breeding groups consisting of two generations we were able to identify several DQ haplotypes by segregation analysis using the previously developed typing protocol for DQA1.
截至目前，恒河猴DQB1基因座的19个等位基因序列已被公布。参照这些序列，我们开发了一种Mamu-DQB1等位基因分型方案，该方案通过额外的克隆、序列分析和分离研究得到了验证。该方案基于使用一对特异性引物扩增第二外显子，随后用多达10种不同的限制性内切酶消化PCR产物。由于大多数扩增的第二外显子序列在用至少一种选定的限制性内切酶消化后会产生独特的图谱，因此可以在纯合和杂合组合中鉴定等位基因。通过使用该方案，我们分析了182只恒河猴的DNA样本。在这些样本中检测到两个新的Mamu-DQB1等位基因，随后进行了克隆并确定了它们的核酸序列。由于我们对由两代组成的四个完整繁殖群体进行了分型，因此能够使用先前开发的DQA1分型方案通过分离分析鉴定几种DQ单倍型。
