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基于序列和聚合酶链反应-序列特异性引物法对贵州小型猪猪白细胞抗原I类基因的分子特征分析

Molecular characterization of swine leukocyte antigen class I genes by sequence-based and PCR-SSP method in Guizhou minipigs.

作者信息

Xia Jihan, Liu Zizhan, Xin Leilei, Wang Zaigui, Qian Lin, Wu Shuguang, Yang Shulin, Li Kui

机构信息

Institute of Animal Science, Chinese Academy of Agricultural Sciences, NO.2 Yuanmingyuan West Road, Beijing, 100193, People's Republic of China.

出版信息

Mol Biol Rep. 2014 Dec;41(12):7775-82. doi: 10.1007/s11033-014-3670-z. Epub 2014 Aug 6.

Abstract

The highly polymorphic swine leucocyte antigen (SLA) genes play an important role in swine immune responses to infectious diseases, vaccines and production performance. The pig resource with well defined SLA genes is useful for xenotransplantation and immunological studies. In this study, we have characterized three SLA class I genes (SLA-1, SLA-3, SLA-2) of 22 founder Guizhou minipigs using sequence-based typing method. Thirteen alleles were detected in this population, compared with the SLA allele sequences in GenBank, 11 of 13 SLA class I alleles were novel in Guizhou minipigs. There are four SLA I haplotypes, none of them previously reported in other pigs. Based on these alleles sequences information, we developed a simple method implemented to SLA-typing for unknown offsprings of Guizhou minipigs, relying on designed 13 sequence specific primers that could discriminate each one among which located in each locus using PCR in a SLA typing assay. According the combination methods of sequence-based typing and PCR-SSP, we were able to rapidly conduct SLA typing for Guizhou breeding stock and identify four SLA haplotypes present in the herd. This resource population of SLA-defined Guizhou minipigs will be useful as animal models for xenotransplantation and further immunological research.

摘要

高度多态的猪白细胞抗原(SLA)基因在猪对传染病、疫苗的免疫反应以及生产性能方面发挥着重要作用。具有明确SLA基因的猪资源对于异种移植和免疫学研究很有用。在本研究中,我们使用基于序列的分型方法对22头贵州小型猪的奠基者的三个SLA I类基因(SLA-1、SLA-3、SLA-2)进行了特征分析。在该群体中检测到13个等位基因,与GenBank中的SLA等位基因序列相比,13个SLA I类等位基因中有11个在贵州小型猪中是新的。有四种SLA I单倍型,之前在其他猪中均未报道过。基于这些等位基因序列信息,我们开发了一种简单的方法,用于对贵州小型猪的未知后代进行SLA分型,该方法依赖于设计的13条序列特异性引物,这些引物可以在SLA分型检测中通过PCR区分位于每个位点的等位基因。根据基于序列的分型和PCR-SSP的组合方法,我们能够快速对贵州种猪进行SLA分型,并鉴定出该群体中存在的四种SLA单倍型。这种SLA定义明确的贵州小型猪资源群体将作为异种移植和进一步免疫学研究的动物模型。

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