Kobori J A, Winoto A, McNicholas J, Hood L
Division of Biology, California Institute of Technology, Pasadena 91125.
J Mol Cell Immunol. 1984;1(2):125-35.
Using Southern DNA hybridization techniques, restriction enzyme site polymorphisms have been used to correlate the molecular maps of the murine major histocompatibility complex (MHC) I region with the genetic map derived from analyses of recombinant mouse strains. The data indicated that the DNA that maps between the I-A and I-E subregions is limited to 3.4 kilobases (kb) and includes the 3' end of the E beta gene. According to classical genetic mapping by recombinational analysis of serological markers, this region should encode the I-B and I-J subregions. These observations are surprising in two respects. First, 3.4 kb is a small amount of DNA to encode even one complete murine gene. Second, this region, which putatively encodes the I-J gene, appears to reside at least partially within the E beta gene. To analyze these apparent paradoxes, further, we cloned the 3.4-kb region in question from six I-region combinant strains [B10.A(3R), B10.a(5R), B10.A(4R), B10.GD, B10.HTT, and B10.S(9R)] and four strains used in the derivation of the recombinants (B10.D2, B10.A, C57BL/10, and ASW) into a lambda phage vector. By direct restriction enzyme mapping of polymorphic sites, we have confirmed the previously identified boundaries of the I-A and I-E subregions and have narrowed the estimate of the distance between these subregions to approximately 2.0 kb of DNA. This 2.0-kb region encompasses part of the intron between the first- (beta 1) and second-domain (beta 2) exons and the second-domain exon (beta 2) of the E beta gene.(ABSTRACT TRUNCATED AT 250 WORDS)
利用Southern DNA杂交技术,限制性酶切位点多态性已被用于将小鼠主要组织相容性复合体(MHC)I区的分子图谱与通过重组小鼠品系分析得出的遗传图谱相关联。数据表明,定位在I-A和I-E亚区之间的DNA长度限制在3.4千碱基(kb),并且包括Eβ基因的3'端。根据通过血清学标记的重组分析进行的经典遗传图谱绘制,该区域应该编码I-B和I-J亚区。这些观察结果在两个方面令人惊讶。首先,3.4 kb的DNA量即使要编码一个完整的小鼠基因都显得很少。其次,这个推测编码I-J基因的区域似乎至少部分位于Eβ基因内。为了进一步分析这些明显的矛盾之处,我们从六个I区组合品系[B10.A(3R)、B10.a(5R)、B10.A(4R)、B10.GD、B10.HTT和B10.S(9R)]以及用于构建重组体的四个品系(B10.D2、B10.A、C57BL/10和ASW)中克隆了上述3.4 kb区域,并将其插入λ噬菌体载体。通过对多态性位点进行直接限制性酶切图谱分析,我们确认了先前确定的I-A和I-E亚区的边界,并将这两个亚区之间的距离估计缩小到约2.0 kb的DNA。这个2.0 kb区域包括Eβ基因第一结构域(β1)和第二结构域(β2)外显子之间的部分内含子以及第二结构域外显子(β2)。(摘要截短至250字)