牦牛RGS1基因克隆及其在发情周期卵巢中的表达

发布时间：2018-01-27 03:08

本文关键词： 牦牛 RGS 卵巢克隆表达　出处：《畜牧兽医学报》2017年02期 　论文类型：期刊论文

【摘要】：本研究旨在阐明牦牛RGS1基因的表达特性及其在牦牛发情周期中卵巢组织的表达。通过采集不同发情周期的牦牛卵巢及肌肉、脾、心、肾、胃、小肠、小脑、肝和肺组织,以GenBank上已发布的黄牛RGS1基因序列设计引物,利用RT-PCR方法扩增克隆牦牛RGS1基因及检测其在牦牛各组织中的表达谱,并使用相关生物信息学软件分析RGS1基因的结构和功能。采用实时荧光定量PCR法检测RGS1基因在牦牛发情周期卵巢中mRNA的表达水平。结果表明,本试验克隆得到牦牛RGS1基因718bp的cDNA序列。同源性与进化树分析表明,牦牛RGS1核苷酸序列与黄牛、野牦牛等大多数哺乳动物的遗传距离较近,其在进化进程中相对较保守。牦牛RGS1基因CDS区为591bp,编码196个氨基酸残基,相对分子质量为22.48ku,不含跨膜结构和信号肽,为亲水性蛋白和非分泌蛋白。蛋白二级结构和三级结构以α-螺旋和自由卷曲结构为主。RGS1基因在所选牦牛组织样品中均有表达,其中在小肠、小脑、心和胃中表达量最高。荧光定量PCR结果显示,牦牛黄体期卵巢中RGS1mRNA的表达量极显著高于卵泡期和红体期(P0.01),卵泡期中RGS1mRNA表达水平高于红体期,但差异不显著(P0.05)。本研究成功克隆了RGS1基因及其在牦牛卵巢3个时期中mRNA的差异性,表明该基因参与发情周期卵巢的内分泌活动调控。
[Abstract]:The purpose of this study was to elucidate the expression characteristics of yak RGS1 gene and its expression in yak oestrous cycle. The yak ovary, muscle, spleen, heart, kidney, stomach and small intestine were collected from different estrous cycles. The primer was designed for cerebellar, liver and lung tissues based on the RGS1 gene sequence of cattle published on GenBank. RT-PCR method was used to amplify and clone yak RGS1 gene and to detect its expression profile in yak tissues. The structure and function of RGS1 gene were analyzed by bioinformatics software. The expression of RGS1 gene in ovarium of yak estrous cycle was detected by real-time fluorescence quantitative PCR. Ming. In this study, 718bp cDNA sequence of yak RGS1 gene was cloned. Homology and phylogenetic tree analysis showed that yak RGS1 nucleotide sequence was similar to that of yellow cattle. The genetic distance of most mammals, such as wild yak, is close, and its evolution is relatively conservative. The CDS region of yak RGS1 gene is 591 BP, encoding 196 amino acid residues. The relative molecular weight is 22.48ku.There is no transmembrane structure or signal peptide. The protein secondary structure and tertiary structure are 伪 -helix and free curl structure. RGS1 gene is expressed in selected yak tissue samples, including small intestine and cerebellum. The results of fluorescence quantitative PCR showed that the expression of RGS1mRNA in ovaries in luteal phase of yak was significantly higher than that in follicular phase and erythrosomal phase (P0.01). The expression of RGS1mRNA in follicular phase was higher than that in erythroid phase. But the difference was not significant (P 0.05). The RGS1 gene and the difference of mRNA in three stages of yak ovary were cloned successfully in this study. It is suggested that the gene is involved in the regulation of endocrine activity in the ovarium of estrous cycle.
【作者单位】：西南民族大学生命科学与技术学院;西南民族大学青藏高原研究院;
【基金】：四川省科技支撑计划(2014NZ0114) 西南民族大学中央高校基本科研业务费专项资金(2016NZYQN38)
【分类号】：S823.85
【正文快照】： G蛋白信号转导调节因子(Regulator of G pro-tein signaling,RGS)是机体内负责协调对各种外界复杂刺激信号而产生的生理反应的分子开关[1],最早由H.G.Dohlman等在啤酒酵母中发现[2],后被证实在哺乳动物中也存在此蛋白或有相同结构和功能的物质。RGS其与细胞表面偶联受体(GPCRs

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