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首页> 外文期刊>Tissue antigens. >Analysis of the complete cDNA sequences of HLA-DRB1 alleles with group-specific amplification primers in the Chinese Han population.
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Analysis of the complete cDNA sequences of HLA-DRB1 alleles with group-specific amplification primers in the Chinese Han population.

机译：用中国汉族人群的组特异性扩增引物分析HLA-DRB1等位基因的完整cDNA序列。

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摘要

Currently for the majority of HLA-DRB1 alleles the focus has been mainly on exon 2 and complete cDNA sequences of HLA-DRB1 alleles are rare. In this study, we analyzed the complete coding sequences of partial alleles of HLA-DRB1 locus. The cDNA was amplified by polymerase chain reaction using the group-specific primers located in the 5'- and 3'-untranslated regions to obtain the complete coding sequences. The amplification products were sequenced using an ABI BigDye(R) Terminator Cycle Sequencing kit. The HLA-DRB1 allele phylogenetic tree was analyzed by dnaman software. Full-length cDNA sequences of 22 HLA-DRB1 alleles were obtained in this study. HLA-DRB1*08:09, DRB1 *12:02:01, and DRB1*13:12 alleles were first reported for complete coding sequences. The sequences of exon 1 of HLA-DRB1*04:06:01, DRB1*08:03:02, and DRB1 *14:07:01 were newly presented. The complete coding sequences of HLA-DRB1 *01:01:01, DRB1*03:01:01:01, DRB1*04:01:01, DRB1*04:05:01, DRB1*07:01:01: 01, DRB1*09:01:02, DRB1*10:01:01, DRB1*11:01:01, DRB1*12:01:01, DRB1*13: 01:01, DRB1*13:02:01, DRB1*14:04, DRB1*14:54, DRB1*15:01:01:01, DRB1*15: 02:01, and DRB1*16:02:01 were identical to those previously reported. Forty polymorphic positions in complete coding sequences outside exon 2 of these HLA-DRB1 alleles were confirmed. According to the phylogenetic tree of full-length coding sequence, the HLA-DRB1 allele was classified into seven major allelic lineages. In conclusion, a protocol for HLA-DRB1 cDNA amplification and sequencing was improved and the data may help to determine the polymorphism of coding sequences outside exon 2.
机译：目前，对于大多数HLA-DRB1等位基因，重点主要放在外显子2上，而HLA-DRB1等位基因的完整cDNA序列却很少。在这项研究中，我们分析了HLA-DRB1基因座的部分等位基因的完整编码序列。通过使用位于5'-和3'-非翻译区的组特异性引物，通过聚合酶链反应扩增cDNA，以获得完整的编码序列。使用ABI BigDye终止子循环测序试剂盒对扩增产物进行测序。通过dnaman软件分析了HLA-DRB1等位基因系统树。在这项研究中获得了22个HLA-DRB1等位基因的全长cDNA序列。首先报道HLA-DRB1 * 08：09，DRB1 * 12：02：01和DRB1 * 13：12等位基因的完整编码序列。新提出了HLA-DRB1 * 04：06：01，DRB1 * 08：03：02和DRB1 * 14：07：01外显子1的序列。 HLA-DRB1 * 01：01：01，DRB1 * 03：01：01：01，DRB1 * 04：01：01，DRB1 * 04：05：01，DRB1 * 07：01：01：01的完整编码序列，DRB1 * 09：01：02，DRB1 * 10：01：01，DRB1 * 11：01：01，DRB1 * 12：01：01，DRB1 * 13：01：01，DRB1 * 13：02：01，DRB1 * 14：04，DRB1 * 14：54，DRB1 * 15：01：01：01，DRB1 * 15：02：01和DRB1 * 16：02：01与先前报告的相同。在这些HLA-DRB1等位基因的外显子2外的完整编码序列中，确定了40个多态位点。根据全长编码序列的系统树，HLA-DRB1等位基因被分为七个主要等位基因谱系。总之，改进了用于HLA-DRB1 cDNA扩增和测序的协议，该数据可能有助于确定外显子2外部编码序列的多态性。

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