Analysis of HLA-DP allelic sequence polymorphism using the in vitro enzymatic DNA amplification of DP-alpha and DP-beta loci.
Analysis of HLA-DP allelic sequence polymorphism using the in vitro enzymatic DNA amplification of DP-alpha and DP-beta loci.
复制标题
使用 DP-α 和 DP-β 位点的体外酶促 DNA 扩增分析 HLA-DP 等位基因序列多态性。
DOI：
10.1007/978-3-662-39946-0_119
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发表时间：
1988
影响因子：
4.4
通讯作者：
H. Erlich
中科院分区：
文献类型：
--
作者：
T. Bugawan;G. Horn;C. M. Long;E. Mickelson;J. Hansen;G. Ferrara;G. Angelini;H. Erlich
在一组34个DP型细胞系中分析了HLA DP-α和DP-β基因的等位基因序列变异。这些基因的多态性的第二个外显子特异性扩增在体外的聚合酶链反应方法，使用耐热DNA聚合酶的栖热菌，水生。对含有扩增的DP-β序列的M13克隆的分析揭示了总共14个等位基因变体。一般来说，特定的等位基因DP-β序列与每种定义的DPw 1-w 6类型相关，β等位基因亚型显示为DPw 2和DPw 4特异性。还鉴定了与任何T细胞定义的特异性（DP“空白”）不相关的另外六个DP-β等位基因。仅检测到先前表征的DP-α的两个等位基因。这些观察结果表明T细胞定义的DP特异性由β链上的多态性残基决定。DP-β中的序列多态性聚集在几个特定区域中，并且可以使用序列特异性寡核苷酸探针和聚合酶链反应扩增的DNA以快速斑点印迹格式进行检测。这种方法提供了一种简单且信息丰富的DP分型方法。将来自四名DP型乳糜泻患者的DP-β序列与对照个体中DP-β等位基因的分布进行比较。
Allelic sequence variation of the HLA DP-alpha and DP-beta genes has been analyzed in a panel of 34 DP-typed cell lines. The polymorphic second exon of these genes was specifically amplified in vitro by the polymerase chain reaction method, using the thermostable DNA polymerase of Thermus, aquaticus. The analysis of M13 clones containing the amplified DP-beta sequences revealed a total of 14 allelic variants. In general, specific allelic DP-beta sequences were associated with each of the defined DPw1-w6 types, with beta allele subtypes revealed for the DPw2 and DPw4 specificities. An additional six DP-beta alleles which did not correlate with any of the T cell-defined specificities (DP "blanks") were also identified. Only the two previously characterized alleles of DP-alpha were detected. These observations suggest that the T cell-defined DP specificities are determined by polymorphic residues on the beta-chain. The sequence polymorphisms in DP-beta are clustered in a few specific regions, and can be detected using sequence-specific oligonucleotide probes and polymerase chain reaction amplified DNA in a rapid dot-blot format. This approach provides a simple and informative method of DP typing. The DP-beta sequences derived from four DP-typed celiac disease patients were compared with the distribution of DP-beta alleles in control individuals.