本文選題：豬白細(xì)胞抗原DO座位alpha亞基　切入點(diǎn)：版納微型豬近交系　出處：《中國(guó)免疫學(xué)雜志》2017年06期 　論文類型：期刊論文

【摘要】：目的:克隆版納微型豬近交系(Banna mini-pig inbred line,BMI)白細(xì)胞抗原DO座位alpha亞基(SLA-DOA)的編碼序列,并對(duì)其19個(gè)重要組織mRNA表達(dá)作半定量分析。方法:RT-PCR方法獲得SLA-DOA的全長(zhǎng)cDNA序列;半定量RTPCR方法分析其在BMI重要組織的mRNA表達(dá)譜,并運(yùn)用生物信息學(xué)對(duì)其核酸及蛋白序列進(jìn)行分析,構(gòu)建系統(tǒng)進(jìn)化樹(shù)。結(jié)果:BMI SLA-DOA cDNA長(zhǎng)度為1 079 bp,編碼250個(gè)氨基酸,蛋白質(zhì)分子量為27.81 k D,等電點(diǎn)為6.48;位于豬7號(hào)染色體,由4個(gè)外顯子和3個(gè)內(nèi)含子構(gòu)成;其mRNA高表達(dá)于淋巴結(jié)和胃,低表達(dá)于心臟、皮膚和十二指腸,在其他14種組織中不表達(dá);SLA-DOA蛋白存在1個(gè)信號(hào)肽,1個(gè)跨膜螺旋,3個(gè)保守域,4個(gè)O連接的糖基化位點(diǎn),18個(gè)磷酸化位點(diǎn),位于細(xì)胞質(zhì)的概率是94.1%;系統(tǒng)進(jìn)化分析表明,在所選擇的6個(gè)物種中,BMI(豬)與牛的親緣關(guān)系最近。結(jié)論:本研究成功克隆了版納微型豬近交系SLA-DOA基因并進(jìn)行了生物信息學(xué)功能分析和多種組織表達(dá)分析,為版納微型豬近交系的異種器官移植研究奠定基礎(chǔ)。
[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.
【作者單位】： 云南農(nóng)業(yè)大學(xué)云南省版納微型豬近交系重點(diǎn)實(shí)驗(yàn)室;云南農(nóng)業(yè)大學(xué)農(nóng)科專業(yè)基礎(chǔ)實(shí)驗(yàn)教學(xué)示范中心;云南農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)技術(shù)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31160439,31660650,31660637,31460580) 云南省應(yīng)用基礎(chǔ)研究計(jì)劃項(xiàng)目(2016FB046,2016FD027)資助
【分類號(hào)】：Q78;R617

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