水牛FADS2基因的电子克隆及序列分析

发布时间：2018-05-04 22:01

  本文选题：水牛 + FADS基因　； 参考：《中国畜牧兽医》2017年10期

【摘要】：试验旨在利用电子克隆法对水牛Δ6脂肪酸脱氢酶(Δ6-fatty acid desaturases,FADS2)基因进行克隆和生物信息学分析,为探究FADS2基因对水牛泌乳性能的作用机制奠定基础。以奶牛FADS2基因序列(Gen Bank登录号:NM_001083444.1)为探针设计引物,利用电子克隆法克隆水牛FADS2基因,并通过RT-PCR验证,对FADS2基因的序列特征进行生物信息学分析。测序结果表明,水牛FADS2基因序列全长为36 600 bp,由12个外显子和11个内含子组成,包含一个长1 335 bp的开放阅读框,可编码444个氨基酸。序列同源性分析显示,水牛FADS2基因编码序列与牦牛、黄牛、人、猪、家兔、虎鲸和褐家鼠序列的同源性分别为98.88%、98.88%、89.66%、90.79%、90.85%、92.35%和87.11%。蛋白质预测分析表明,水牛FADS2蛋白分子质量为52.51 ku,理论等电点(p I)为8.75,呈弱碱性,属于亲水性蛋白,无信号肽。系统进化树分析结果表明,FADS2基因在不同物种及进化的过程中具有高度保守性,其中水牛与牦牛、黄牛亲缘关系较近,与褐家鼠亲缘关系较远。水牛FADS2基因的成功克隆为今后阐明水牛泌乳性能的作用机制奠定了基础。
[Abstract]:The aim of this study was to clone the 螖 6 fatty acid dehydrogenase (螖 6-fatty acid desaturus FADS2) gene from buffalo by electronic cloning and to analyze its bioinformatics, so as to lay a foundation for exploring the mechanism of FADS2 gene acting on buffalo lactation performance. FADS2 gene sequence Gen Bank accession number: NM0083444.1) was used as a probe to design primers to clone buffalo FADS2 gene by electronic cloning, and the sequence characteristics of FADS2 gene were analyzed by bioinformatics through RT-PCR verification. The sequencing results showed that the buffalo FADS2 gene was 36,600 BP in length and consisted of 12 exons and 11 introns, which contained an open reading frame of 1 335 BP and encoded 444 amino acids. Sequence homology analysis showed that the homology of buffalo FADS2 gene coding sequence with yak, yellow cattle, human, pig, rabbit, orcas and brown rat sequences was 98.8888, 98.888.88 and 89.66, respectively. The homology of buffalo FADS2 gene coding sequence was 90.85% and 92.35% and 87.11% respectively with yak, yellow cattle, human being, rabbit and Rattus norvegicus. Protein prediction and analysis showed that the molecular weight of buffalo FADS2 protein was 52.51 kuand the theoretical isoelectric point (Ip) was 8.75, which was weakly alkaline and belonged to hydrophilic protein with no signal peptide. Phylogenetic tree analysis showed that the FADS2 gene was highly conserved in different species and evolution, among which buffalo was closely related to yak and yellow cattle, and far related to Rattus norvegicus (Rattus norvegicus). The successful cloning of buffalo FADS2 gene laid a foundation for elucidating the mechanism of buffalo lactation.
【作者单位】： 中国农业科学院广西水牛研究所 农业部(广西)水牛遗传繁育重点实验室;
【基金】：广西科技重大专项(桂科AA16450002) 广西科技公关国合项目(桂科合1504001-3) 广西自然科学基金青年项目(2015GXNSFBA139103) 广西水产畜牧科技推广应用项目(桂渔牧科201633009)
【分类号】：S823.83
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本文编号：1844843