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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。
Krüppel-like factors (KLFs) are a group of zinc finger proteins that are closely related to the transcriptional regulation of eukaryotic genes. The purpose of this study was to clone the CDS region of KLF13 gene from Capra hircus and to detect its expression in different tissues of goat and to identify the expression pattern of it during cell differentiation. Four randomly selected 24-month-old healthy Jane sheep were randomly selected and hearts, liver, spleen, lung, kidney, subcutaneous fat, dorsal longissimus, biceps femoris and triceps brachii were sampled after slaughter The total RNA of the tissue was extracted and the sequence of KLF13 was cloned by RT-PCR. Bioinformatics analysis was performed. The expression of KLF13 mRNA was detected by real-time quantitative polymerase chain reaction (qRT-PCR) 7 d) and myoblasts (0, 2, 3, 4, 5 d) induced KLF13 m RNA expression. The results showed that the cloned KLF13 c DNA sequence was 991 bp in length (Gen Bank accession number: KU041755), which contained 876 bp in CDS region, 12 bp in 5’UTR sequence and 103 bp in 3’UTR sequence, encoding 292 amino acid residues. The similarity of the deduced amino acid sequence of Goat KLF13 gene to Ovis aries, Bos taurus, Sus scrofa and Mus musculus was 100%, 97.9%, 96.6% and 90.8% respectively. The goat KLF13 gene has the closest genetic relationship with sheep, followed by Homo sapiens, pigs and mice. KLF13 m RNA has the highest level of expression in goat spleen and lungs, but lower in muscle. KLF13m RNA expression gradually decreased during the differentiation of precursor adipocytes, but gradually increased during myogenic differentiation (except for 5 days). The results suggest that KLF13 gene may play an important role in the regulation of goat precursor adipocyte differentiation. This experiment provides the basic information for studying the role of KLF13 gene in goat precursor adipocyte differentiation.