【摘要】：試驗(yàn)旨在克隆北極狐及烏蘇里貉抑制素α(inhibinα,INHα)亞基基因并對其進(jìn)行生物信息學(xué)分析。根據(jù)GenBank中犬科INHα預(yù)測mRNA序列(登錄號:XM_545660.5)設(shè)計(jì)1對引物,用RT-PCR技術(shù)從北極狐及烏蘇里貉的卵巢組織中擴(kuò)增出INHα亞基基因,同時(shí)將其插入到克隆載體中,進(jìn)行測序及生物信息學(xué)分析。測序結(jié)果表明,北極狐及烏蘇里貉的INHα亞基基因CDS序列全長為1 107bp,編碼369個(gè)氨基酸。北極狐及烏蘇里貉的INHα亞基基因與犬的同源性最高,分別為97.9%與97.6%。系統(tǒng)進(jìn)化樹分析表明,北極狐及烏蘇里貉與犬親緣關(guān)系較近,同時(shí)也說明INHα亞基基因在不同物種及進(jìn)化過程中具有高度保守性。對INHα亞基蛋白的高級結(jié)構(gòu)預(yù)測發(fā)現(xiàn),由于半胱氨酸間形成的二硫鍵導(dǎo)致其采用"蝴蝶形"或"開放手"構(gòu)型,其中α-螺旋形成分子的"手腕"結(jié)構(gòu),β-折疊形成分子的"手指"結(jié)構(gòu)。本研究成功克隆了北極狐及烏蘇里貉的INHα亞基基因,同時(shí)進(jìn)行了系統(tǒng)的生物信息學(xué)分析,為今后研究抑制素在卵母細(xì)胞-顆粒細(xì)胞同步發(fā)育過程中的生物學(xué)功能奠定了基礎(chǔ)。
[Abstract]:The aim of this study was to clone inhibin 偽 (INH 偽) subunit genes from arctic fox and raccoon dog and to analyze them by bioinformatics. According to the INH 偽 predicted mRNA sequence (accession number: XM_545660.5) in GenBank, a pair of primers were designed. The INH 偽 subunit gene was amplified from the ovarian tissues of Arctic fox and raccoon dog by RT-PCR technique and inserted into the clone vector. Sequencing and bioinformatics analysis were performed. The results of sequencing showed that the CDS sequence of INH 偽 subunit gene of arctic fox and raccoon was 1 107 BP, encoding 369 amino acids. The homology of INH 偽 subunit gene between arctic fox and raccoon dog was 97.9% and 97.6%, respectively. Phylogenetic tree analysis showed that arctic fox and raccoon dog were closely related to dogs, and that INH 偽 subunit gene was highly conserved in different species and evolution process. The advanced structure prediction of INH 偽 subunit protein showed that the disulfide bond formed between cysteine resulted in the "butterfly" or "open hand" configuration, in which 偽 -helix formed the "wrist" structure of the molecule. 尾-fold to form the "finger" structure of the molecule. In this study, the INH 偽 subunit genes of arctic fox and raccoon dog were cloned successfully, and the bioinformatics analysis was carried out systematically, which laid a foundation for studying the biological function of inhibin in the process of oocyte granulosa cell development in the future.
【作者單位】： 中國農(nóng)業(yè)科學(xué)院特產(chǎn)研究所;中國農(nóng)業(yè)科學(xué)院特種經(jīng)濟(jì)動(dòng)物分子生物學(xué)重點(diǎn)實(shí)驗(yàn)室;
【基金】：中國農(nóng)業(yè)科學(xué)院創(chuàng)新工程(特種經(jīng)濟(jì)動(dòng)物繁殖創(chuàng)新團(tuán)隊(duì));中國農(nóng)業(yè)科學(xué)院基本科研業(yè)務(wù)費(fèi)專項(xiàng)(1610342017025) 吉林省國際合作項(xiàng)目(20170414049GH) 國家自然基金面上項(xiàng)目(31772606)
【分類號】：S865.2

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