Zhou J H, Ferencik S, Rebmann V, Yang D L, Lu M, Roggendorf M, Grosse-Wilde H
Institute of Immunology, Institute of Virology, University Hospital of Essen, Virchowstrasse 171, 45122 Essen, Germany.
Tissue Antigens. 2003 Mar;61(3):240-8. doi: 10.1034/j.1399-0039.2003.00036.x.
The woodchuck (Marmota monax) is an animal model that is used in the study of human hepatitis B virus ( HBV ) infection. A knowledge of woodchuck MHC class I (Mamo-I) genes and gene products is therefore essential for understanding the antigen-specific T-cell responses in this animal model. A number of Mamo-I genes have been identified by molecular cloning and sequencing. However, the allelic nature of these genes has not been proven by classical genetics like the segregation analysis in families. In this study, we analyzed the allelic diversity of Mamo-I in two three-generation woodchuck families including 15 members by sequencing of Mamo-I genes and immunoblotting of Mamo-I proteins after one-dimensional isoelectric focusing (1D-IEF). In addition to four published Mamo-I alleles, six new alleles that belonged to the same locus as the known Mamo-I alleles (Mamo-A) were found within the two woodchuck families. A typical Mendelian segregation of Mamo-I gene and antigens was observed in the families studied. For simple and rapid detection of allelic variability of Mamo-I gene, a typing method based on the detection of PCR products amplified by sequence specific primers (SSP) has been developed and tested in 41 unrelated animals. The most prevalent allele was Mamo-A01 with a frequency of 21.9% followed by Mamo-A07 (12.2%). Our study established Mamo-A as a classical MHC class I locus by the polymorphic and allelic nature of Mamo-I gene in the woodchuck.
土拨鼠（Marmota monax）是用于人类乙型肝炎病毒（HBV）感染研究的动物模型。因此，了解土拨鼠MHC I类（Mamo-I）基因和基因产物对于理解该动物模型中的抗原特异性T细胞反应至关重要。通过分子克隆和测序已鉴定出多个Mamo-I基因。然而，这些基因的等位基因性质尚未通过经典遗传学方法（如家族中的分离分析）得到证实。在本研究中，我们通过对Mamo-I基因进行测序以及在一维等电聚焦（1D-IEF）后对Mamo-I蛋白进行免疫印迹分析，对两个包含15个成员的三代土拨鼠家族中的Mamo-I等位基因多样性进行了分析。除了四个已发表的Mamo-I等位基因外，在这两个土拨鼠家族中还发现了六个与已知Mamo-I等位基因（Mamo-A）属于同一基因座的新等位基因。在所研究的家族中观察到了Mamo-I基因和抗原的典型孟德尔分离现象。为了简单快速地检测Mamo-I基因的等位基因变异性，我们开发了一种基于检测序列特异性引物（SSP）扩增的PCR产物的分型方法，并在41只无关动物中进行了测试。最常见的等位基因是Mamo-A01，频率为21.9%，其次是Mamo-A07（12.2%）。我们的研究通过土拨鼠中Mamo-I基因的多态性和等位基因性质将Mamo-A确立为一个经典的MHC I类基因座。
