本文選題：水牛　切入點(diǎn)：波形蛋白(VIM)　出處：《南方農(nóng)業(yè)學(xué)報(bào)》2017年12期 　論文類型：期刊論文

【摘要】：【目的】克隆水牛波形蛋白(VIM)基因,并明確VIM基因在水牛成熟卵母細(xì)胞(MII期)和早期胚胎(2-細(xì)胞階段)中的表達(dá)情況,為研究VIM在卵母細(xì)胞成熟及附植前胚胎發(fā)育中的作用機(jī)理打下基礎(chǔ)。【方法】根據(jù)GenBank上已公布的水牛VIM基因mRNA序列設(shè)計(jì)引物,采用RT-PCR克隆水牛VIM基因,利用在線軟件程序?qū)寺～@得的水牛VIM基因進(jìn)行生物信息學(xué)分析,同時(shí)以細(xì)胞免疫熒光試驗(yàn)檢測(cè)VIM在水牛成熟卵母細(xì)胞(MII期)及早期胚胎(2-細(xì)胞階段)中的表達(dá)情況。【結(jié)果】水牛VIM基因長(zhǎng)度為1469 bp,其編碼區(qū)長(zhǎng)度1401 bp,編碼466個(gè)氨基酸。水牛VIM基因序列與人類、黑猩猩、食蟹猴、家犬、馬和牛的VIM基因序列具有高度的同源性,分別為93%、93%、93%、94%、94%和99%;基于VIM基因序列構(gòu)建的系統(tǒng)發(fā)育進(jìn)化樹也顯示水牛與牛的親緣關(guān)系最近。水牛VIM的分子量為53.73 k D,理論等電點(diǎn)(p I)5.05,主要在線粒體中表達(dá),穩(wěn)定性差(不穩(wěn)定系數(shù)53.65),具有較強(qiáng)的親水性,無(wú)跨膜結(jié)構(gòu),不屬于分泌型蛋白,其蛋白結(jié)構(gòu)以α-螺旋為主。VIM在水牛成熟卵母細(xì)胞(MII期)及早期胚胎(2-細(xì)胞階段)中均有表達(dá),且在成熟卵母細(xì)胞中的表達(dá)量明顯高于早期胚胎�！窘Y(jié)論】水牛VIM基因編碼序列具有較高的保守性,且在水牛成熟卵母細(xì)胞(MII期)中的表達(dá)量明顯高于早期胚胎(2-細(xì)胞階段)。
[Abstract]:[objective] to clone buffalo vimentin (VIM) gene, and to identify the expression of VIM gene in matured buffalo oocytes (MII) and in the early embryo (2-cell stage). In order to study the mechanism of VIM in oocyte maturation and preimplantation embryo development, a primer was designed according to the mRNA sequence of buffalo VIM gene published on GenBank, and RT-PCR was used to clone VIM gene of buffalo. The cloned buffalo VIM gene was analyzed by bioinformatics using online software program. Meanwhile, the expression of VIM in buffalo matured oocytes (MII) and in the early stage of embryo (2-cell) was detected by cellular immunofluorescence assay. [results] the length of buffalo VIM gene was 1469 BP, and the length of coding region was 1401 BP, encoding 466 ammonia. Basic acid, buffalo VIM gene sequence and human, The VIM gene sequence of chimpanzee, crab monkey, domestic dog, horse and cow is highly homologous. The phylogenetic tree based on VIM gene sequence also showed the closest relationship between buffalo and cattle. The molecular weight of buffalo VIM was 53.73 KD, and the theoretical isoelectric point was 5.05, mainly expressed in mitochondria. There was poor stability (unstable coefficient 53.65, strong hydrophilicity, no transmembrane structure and no secretory protein. The protein structure was mainly 偽 -helix in buffalo matured oocytes in MII phase) and in the early embryonic stage of 2-cell). The expression of VIM gene in matured oocytes was significantly higher than that in early embryos. [conclusion] the VIM gene coding sequence of buffalo is highly conserved, and the expression level in matured buffalo oocytes is significantly higher than that in the early embryo 2-cell stage.
【作者單位】： 廣西大學(xué)動(dòng)物繁殖研究所/亞熱帶農(nóng)業(yè)生物資源保護(hù)與利用國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31460603) 廣西研究生教育創(chuàng)新計(jì)劃項(xiàng)目(YCBZ2017005)
【分類號(hào)】：S823.83

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本文編號(hào)：1625709