Elucidation of the evolution of the MHC by genome analysis

为了阐明四个旁系同源区域的基因组织;染色体6p21.3（HLA区域）、1 q21-q25、9 q33-q34和19p13.1-p13.3区域的基因组结构，并了解HLA区域的进化，我们用YAC分析了CD 1基因周围的HLA I类区域、9 q33-q34和1 q22-q23区域的基因组结构，PAC和BAC克隆。通过鸟枪法测定1.8 Mb HLA I类整个区域的基因组序列。在该区域共发现124个基因或候选基因序列和758个微卫星重复序列。在9 q33-q34区域，定位了3个与HLA区域同源的基因，其中HXB -C5 -GRP 78来自YAC克隆的PCR分析，在1 q22-q23区域，用位点特异性引物和STS引物对PCR和PAC克隆的末端区域测序，构建了约1.8Mb的包含5个CD 1基因的YAC和PAC重叠群。10个基因位于CD 1 E的端粒侧，顺序为SPTA 1 - MNDA -FYI - IFI-16 -FCERIA -KRTC 7-CRP-CRPP 1-ARCS - KIAAO 120。此外，从位于KIAA 0120端粒侧的YAC克隆的PCR分析中，定位了12个基因，（ATP1A2 -PXF-CBP-AIM2）- LYAM -D1 S1705和WI-6210之间的（CD 48-LY 9- PPOX）-USF 1-（EAT 2-PBX 1）-ALDH 9-RXRG-POU 2F 1-CD 3 Z-ATAC-（FV-ELAM 1- LNHR）-FMO 1- FASL。结果表明，CD 1基因1 q22-q23区域包含一组具有免疫学意义的基因，如SPTA 1、FY、IFI-16、ECERIA、ARCS、LYAM、CD 48、LY 9、SCM、CD 3 Z、ATAC、ELAM 1、LNHR和FASL。

In order to clarify the gene organization of four paralogous regions ; chromosome 6p21.3 (HLA region), the 1q21-q25, 9q33-q34, and 19p13.1-p13.3 regions, and understanding of the evolution of the HLA region, we have analyzed the genomic structure of the HLA class I region, 9q33-q34 and 1q22-q23 region around the CD1 genes by YAC, PAC and BAC clones. The genomic sequence of the 1.8 megabase (Mb) HLA class I entire region, was determined by the shot gun method. In total, 124 genes or candidate gene sequences and 758 microsatellite repeats were found in this region. In addition, the five pair segments which showed high sequence homology to each other were found in the HLA class I region, suggesting that segmental genome duplication must have occurred during the evolution of the human MHC, ln the 9q33-q34 region, three genes which have homologous genes in the HLA region were mapped, HXB -C5 -GRP78 from PCR analyses of the YAC clones, In the 1q22-q23 region, an approximately 1.8 Mb YAC and PAC contig containing thefive CD1 genes were constructed by PCR using locus-specific and STS primer pairs and sequencing of the end regions of PAC clones. Ten genes were located in the telomeric side of CDlE with the order of SPTA 1 - MNDA -FYI - IFI-16 -FCERIA -KRTC7 -CRP -CRPP1 -ARCS - KIAAO 120 . Further, from PCR analyses of the YAC clones which were located on the telomeric side of KIAA 0120, twelve genes were mapped, (ATP1A2 - PXF - CBP -AIM2)- LYAM - (CD48 - LY9 - PPOX) -USF1 - (EAT2 -PBX1) - ALDH9 - RXRG - POU2F1 -CD3Z - ATAC - (FV- ELAM1 - LNHR) -FMO1 - FASL between D1S1705 and WI-6210. Thus, it was revealed that the 1q22-q23 region around the CD1 genes contained a cluster of genes which had immunological importance, such as SPTA 1, FY, IFI-16, ECERIA, ARCS, LYAM, CD48, LY9, SCM, CD3Z, ATAC, ELAM1, LNHR and FASL .