版纳微型猪近交系SLA-DOA基因克

发布时间：2018-03-03 02:37

  本文选题：猪白细胞抗原DO座位alpha亚基　切入点：版纳微型猪近交系　出处：《中国免疫学杂志》2017年06期 　论文类型：期刊论文

【摘要】：目的:克隆版纳微型猪近交系(Banna mini-pig inbred line,BMI)白细胞抗原DO座位alpha亚基(SLA-DOA)的编码序列,并对其19个重要组织mRNA表达作半定量分析。方法:RT-PCR方法获得SLA-DOA的全长cDNA序列;半定量RTPCR方法分析其在BMI重要组织的mRNA表达谱,并运用生物信息学对其核酸及蛋白序列进行分析,构建系统进化树。结果:BMI SLA-DOA cDNA长度为1 079 bp,编码250个氨基酸,蛋白质分子量为27.81 k D,等电点为6.48;位于猪7号染色体,由4个外显子和3个内含子构成;其mRNA高表达于淋巴结和胃,低表达于心脏、皮肤和十二指肠,在其他14种组织中不表达;SLA-DOA蛋白存在1个信号肽,1个跨膜螺旋,3个保守域,4个O连接的糖基化位点,18个磷酸化位点,位于细胞质的概率是94.1%;系统进化分析表明,在所选择的6个物种中,BMI(猪)与牛的亲缘关系最近。结论:本研究成功克隆了版纳微型猪近交系SLA-DOA基因并进行了生物信息学功能分析和多种组织表达分析,为版纳微型猪近交系的异种器官移植研究奠定基础。
[Abstract]:Objective: to clone the coding sequence of alpha subunit SLA-DOAA of leukocyte antigen do locus of Banna mini-pig inbred lineage BMIA, and to make semi-quantitative analysis of mRNA expression in 19 important tissues of Banna micropig inbred line Banna mini-pig inbred. Methods: the full-length cDNA sequence of SLA-DOA was obtained by RT-PCR. The semi-quantitative RTPCR method was used to analyze the mRNA expression profile in the important tissues of BMI. The nucleic acid and protein sequences were analyzed by bioinformatics, and the phylogenetic tree was constructed. Results the length of SLA-DOA cDNA was 1 079 BP, encoding 250 amino acids. The protein has a molecular weight of 27.81 KD and an isoelectric point of 6.48. It is located on chromosome 7 of pigs and consists of four exons and three introns. Its mRNA is highly expressed in lymph nodes and stomach, and is low in heart, skin and duodenum. There was a signal peptide, a transmembrane helix, three conserved domains, four O-linked glycosylation sites and 18 phosphorylation sites in the other 14 tissues. Among the 6 selected species, BMI (pig) was closely related to cattle. Conclusion: SLA-DOA gene of Banna miniature pig inbred line was cloned successfully, and bioinformatics function analysis and tissue expression analysis were carried out. It lays a foundation for the study of xenotransplantation of Banna miniature pig inbred lines.
【作者单位】： 云南农业大学云南省版纳微型猪近交系重点实验室;云南农业大学农科专业基础实验教学示范中心;云南农业大学动物科学技术学院;
【基金】：国家自然科学基金项目(31160439,31660650,31660637,31460580) 云南省应用基础研究计划项目(2016FB046,2016FD027)资助
【分类号】：Q78;R617

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