Molecular linkage of the HLA-DR, HLA-DQ, and HLA-DO genes in yeast artificial chromosomes.
Molecular linkage of the HLA-DR, HLA-DQ, and HLA-DO genes in yeast artificial chromosomes.
复制标题
酵母人工染色体中 HLA-DR、HLA-DQ 和 HLA-DO 基因的分子连锁。
DOI：
10.1016/0888-7543(91)90065-m
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发表时间：
1991
期刊：
影响因子：
4.4
通讯作者：
Chaplin,DD
中科院分区：
文献类型：
--
作者：
Kozono,H;Bronson,SK;Taillon-Miller,P;Moorti,MK;Jamry,I;Chaplin,DD
从重叠的酵母人工染色体(YAC)克隆中分离到8个主要组织相容性复合体(MHC)II类基因座和新定义的Y3/Ring 4基因座，定义了人类染色体6p21.3的420kb片段。YAC B1D12全长320 kb，包含7个从人类白细胞抗原DRA到人类白细胞抗原DQB2的基因座。一个330kb的YAC，A148A7，从HLADQA1基因座跨越到Y3/Ring 4基因座，延伸了至少130kb的YAC B1D12着丝粒。Southern杂交结果表明，YACB1D12来源于杂合子文库供者的人类白细胞抗原DR3单倍型，YACA148A7来源于杂合子文库供者的人类白细胞抗原DR7单倍型。第三个150kb的YAC，A95C5，位于这个重叠群中，只包含HLA-DRA基因座。通过染色体走行从YAC B1D12的端粒末端分离到第四个300kb的YAC，A76F11。从YAC基因组插入片段末端分离的探针已用于确认克隆之间的重叠。这些分析表明，YAC A76F11的着丝粒末端与YAC A95C5的端粒末端使用相同的基因组EcoRI克隆位点。YAC B1D12仅使用上述EcoRI位点的2.1kb端粒。这些数据表明，在文库建设过程中，某些EcoRI位点被优先使用。这些YAC完成了克隆DNA中人类白细胞抗原复合体的DR和DQ亚区的连接，并为精确分析这部分II类区域提供了底物。
Eight major histocompatibility complex (MHC) class II loci and the newly defined Y3/Ring 4 locus were isolated in overlapping yeast artificial chromosome (YAC) clones defining a 420-kb segment of human chromosome 6p21.3. YAC B1D12 spanning 320 kb contained seven of these loci from HLA-DRA to HLA-DQB2. A 330-kb YAC, A148A7, spanned from the HLA-DQA1 locus through the Y3/Ring 4 locus and extended at least 130 kb centromeric of YAC B1D12. Southern blotting demonstrated that YAC B1D12 derived from the HLA-DR3 haplotype and that YAC A148A7 derived from the HLA-DR7 haplotype of the heterozygous library donor. A third 150-kb YAC, A95C5, lay within this contig and contained only the HLA-DRA locus. A fourth 300-kb YAC, A76F11, was isolated by chromosome walking from the telomeric end of YAC B1D12. Probes isolated from the ends of the YAC genomic inserts have been used to confirm overlaps between the clones. These analyses demonstrated that the centromeric end of YAC A76F11 used the same genomicEcoRI cloning site as the telomeric end of YAC A95C5. YAC B1D12 used anEcoRI site only 2.1 kb telomeric of the aforementionedEcoRI site. These data suggest that certainEcoRI sites are used preferentially during construction of the library. These YACs complete the linkage of the DR and DQ subregions of the HLA complex in cloned DNA and provide the substrate for precise analysis of this portion of the class II region.