Strachan T, Dodge A B, Smillie D, Dyer P A, Sodoyer R, Jordan B R, Harris R
Immunogenetics. 1986;23(2):115-20. doi: 10.1007/BF00377971.
Analysis of available nucleotide sequence data for class I HLA genes has established that the seventh intron is one of the gene regions which expresses the highest degree of locus specificity (the percentage sequence divergence between nonallelic genes minus the percentage sequence divergence between allelic genes). We have subcloned short DNA sequences including this region from the HLA-Cw3 gene. Two clones, pC250 and pC800, were tested by hybridizing them at high stringency to a panel of clones containing class I HLA genes. Under conditions permitting a strong hybridization signal with a C-locus gene, pC800 also expressed a weak but significant hybridization to other class I genes, while pC250 appeared to hybridize exclusively to the C-locus gene. Hybridization of the pC250 probe at high stringency to Hind III-digested genomic DNA from a panel of unrelated individuals and homozygous typing cell lines revealed a single band in all cases. However, equivalent hybridization against Eco RI-digested DNA revealed two hybridization bands, one at 7.9 kb which correlated with the serologically defined Cw5 and Cw8 alleles, and one at 7.6 kb which correlated with the Cw1, Cw2, Cw3, Cw4, Cw6, and Cw7 alleles.
对I类HLA基因现有核苷酸序列数据的分析表明，第七内含子是表达最高程度基因座特异性的基因区域之一（非等位基因之间的序列差异百分比减去等位基因之间的序列差异百分比）。我们从HLA - Cw3基因中克隆了包括该区域的短DNA序列。通过将两个克隆pC250和pC800与一组含有I类HLA基因的克隆进行高严谨度杂交来进行检测。在允许与C基因座基因产生强烈杂交信号的条件下，pC800也与其他I类基因表现出微弱但显著的杂交，而pC250似乎仅与C基因座基因杂交。将pC250探针与来自一组无关个体和纯合分型细胞系的经Hind III消化的基因组DNA进行高严谨度杂交，在所有情况下均显示出一条带。然而，与经Eco RI消化的DNA进行的等效杂交显示出两条杂交带，一条在7.9 kb处，与血清学定义的Cw5和Cw8等位基因相关，另一条在7.6 kb处，与Cw1、Cw2、Cw3、Cw4、Cw6和Cw7等位基因相关。
