山羊KLF13基因的克隆及表达分析

发布时间：2018-05-20 21:08

  本文选题：山羊 + KLF　； 参考：《农业生物技术学报》2017年01期

【摘要】：Krüppel样因子(Krüppel-like factors,KLFs)是一组与真核基因转录调控密切相关的锌指蛋白。本研究旨在克隆山羊(Capra hircus)KLF13基因的CDS区序列,检测其在山羊不同组织中表达水平,并鉴定在细胞分化过程中的表达规律。随机选取24月龄的健康简州大耳羊公羊4只,屠宰后分别采集心、肝、脾、肺、肾、皮下脂肪、背最长肌、股二头肌和臂三头肌组织样品,提取组织总RNA,利用RT-PCR法克隆山羊KLF13基因序列,并进行生物信息学分析;利用实时定量PCR法检测KLF13 m RNA的表达水平,并检测前体脂肪细胞(0、3、5、7 d)和成肌细胞(0、2、3、4、5 d)诱导分化后的KLF13 m RNA表达水平。结果显示,克隆得到KLF13 c DNA序列长991 bp(Gen Bank登陆号:KU041755),其中包含CDS区876 bp,5'UTR序列12 bp,3'UTR序列103 bp,编码292个氨基酸残基。山羊KLF13基因编码的氨基酸序列与绵羊(Ovis aries)、牛(Bos taurus)、猪(Sus scrofa)、小鼠(Mus musculus)的相似性分别为100%、97.9%、96.6%和90.8%。山羊KLF13基因与绵羊具有最近的亲缘关系,其次为牛、人(Homo sapiens)、猪和小鼠。KLF13 m RNA在山羊脾和肺中具有最高的表达水平,而在肌肉中的表达丰度较低。在前体脂肪细胞的分化过程中KLF13m RNA表达水平逐渐降低,而在成肌细胞分化过程中则逐渐上升(除5 d外)。结果提示,KLF13基因可能在调控山羊前体脂肪细胞分化过程中具有重要作用。本实验为研究KLF13基因在山羊前体脂肪细胞分化中的作用提供了基础资料。
[Abstract]:Kr 眉 ppel like factor Kr 眉 ppel-like factors KLFs is a group of zinc finger proteins closely related to the transcriptional regulation of eukaryotic genes. The purpose of this study was to clone the CDS sequence of Capra hircus)KLF13 gene from goat, detect its expression level in different tissues of goat, and identify the expression pattern of capra hircus)KLF13 gene in the process of cell differentiation. Samples of heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi muscle, biceps femoris muscle and triceps arm muscle were collected after slaughter. The total RNAs were extracted, the goat KLF13 gene sequence was cloned by RT-PCR method, and bioinformatics analysis was carried out. The expression level of KLF13 m RNA was detected by real-time quantitative PCR. The expression of KLF13 m RNA in preadipocytes and myoblasts were detected after induction of differentiation for 5 d. The results showed that the length of the KLF13 c DNA sequence was 991 bp(Gen Bank, which contained the CDS region 876 BP ~ (5) ~ (5) UTR sequence (12 BP ~ (3) UTR sequence (103 BP), encoding 292 amino acid residues. The amino acid sequence of goat KLF13 gene was similar to that of Ovis ariesus, Bos taurus, Sus scrofaus and Mus musculus, respectively. The similarity of amino acid sequence of goat KLF13 gene to Ovis ariesus, Bos taurus, Sus scrofaus and Mus musculus was 96. 6% and 98. 8%, respectively. Goat KLF13 gene was closely related to sheep, followed by cattle, Homo sapiensus, pig and mouse. KLF13m RNA had the highest expression level in goat spleen and lung, but lower expression in muscle. During the differentiation of preadipocytes, the expression of KLF13m RNA decreased gradually, but increased gradually during the differentiation of myoblasts (except for 5 days). These results suggest that KLF13 gene may play an important role in regulating the differentiation of goat precursor adipocytes. This study provides basic data for the study of the role of KLF13 gene in the differentiation of goat precursor adipocytes.
【作者单位】： 青藏高原动物遗传资源保护与利用四川省重点实验室;西南民族大学生命科学与技术学院;四川省畜牧科学研究院;
【基金】：中央高校基本科研业务费专项资金项目(No.2016NZYQN36)
【分类号】：S827
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本文编号：1916240