华北农学报 ›› 2021, Vol. 36 ›› Issue (2): 226-232. doi: 10.7668/hbnxb.20191543

所属专题: 畜牧

• 畜牧·水产·兽医 • 上一篇    下一篇

山羊KLF6的分子克隆及其时空表达特征分析

王江林1,2, 王永1, 孟庆勇3, 朱江江1, 林亚秋1,2   

  1. 1. 西南民族大学, 青藏高原动物遗传资源保护与利用教育部、四川省重点实验室, 四川 成都 610041;
    2. 西南民族大学 畜牧兽医学院, 四川 成都 610041;
    3. 中国农业大学 生物学院, 农业生物技术国家重点实验室, 北京 100193
  • 收稿日期:2020-12-17 出版日期:2021-04-28
  • 通讯作者: 林亚秋(1976-),女,内蒙古海拉尔人,教授,博士,主要从事动物遗传育种与繁殖研究。
  • 作者简介:王江林(1994-),男,贵州铜仁人,硕士,主要从事动物遗传育种研究。
  • 基金资助:
    国家自然科学基金项目(31672395);四川省应用基础研究计划重点项目(2018JY0036);西南民族大学中央高校基本科研业务费专项资金(2020PTJS15004)

Molecular Cloning and Spatio-temporal Expression Analysis of KLF6 in Goat

WANG Jianglin1,2, WANG Yong1, MENG Qingyong3, ZHU Jiangjiang1, LIN Yaqiu1,2   

  1. 1. Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Sichuan Province, Southwest Minzu University, Chengdu 610041, China;
    2. College of Animal&Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China;
    3. State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China
  • Received:2020-12-17 Published:2021-04-28

摘要: 旨在获得山羊Kruppel样因子6(Kruppel-like factor 6,KLF6) CDS序列,明确其组织及在皮下脂肪细胞分化过程中的表达模式。利用RT-PCR技术克隆山羊KLF6基因序列,利用生物学软件及在线网站对获得的KLF6基因进行序列分析。利用实时荧光定量PCR(Quantitative Real-time PCR,qPCR)技术检测KLF6在山羊各组织中的表达丰度及成脂诱导分化不同阶段皮下脂肪细胞中的表达水平。结果显示,获得山羊KLF6基因全长序列1 233 bp,其中包括CDS区957 bp,编码318个氨基酸残基,与牛和绵羊的亲缘关系最近。KLF6基因在山羊各个组织中都有广泛表达,且在脂肪组织中高表达,极显著高于其他组织(P<0.01)。KLF6在诱导分化60 h的山羊皮下脂肪细胞中的表达量极显著高于在前体脂肪细胞中的表达水平(P<0.01)。获得山羊KLF6基因序列并明确其分子特征,发现其在脂肪组织中存在高表达,且在分化后表达水平显著高于分化前的表达水平,为最终揭示KLF6调控山羊脂肪细胞分化提供重要数据。

关键词: 山羊, KLF6, 组织表达, 前体脂肪细胞, 细胞分化

Abstract: This study was performed to obtain the sequence of Kruppel-like factor 6(Kruppel-like factor 6, KLF6) CDS, clarify its expression pattern in tissues and subcutaneous adipocyte of goats. The KLF6 gene sequence of goat was cloned by RT-PCR, and the sequence of KLF6 gene was analyzed by biological software and online website. Quantitative real-time PCR (qPCR) was used to detect the expression abundance of KLF6 in various goat tissues and the expression level of subcutaneous adipocytes at different stages of lipogenesis differentiation. The results showed that the full-length sequence of goat KLF6 gene was 1 233 bp, including 957 bp of CDS region, which encoded 318 amino acid residues, and closest relative was the cattle and the sheep. KLF6 gene was widely expressed in various the investigated tissues of goats, the expression level of KLF6 was higher in adipose tissue, which was significantly higher than that in other tissues(P<0.01). The expression of KLF6 in goat subcutaneous adipocytes induced for 60 h was significantly higher than that in preadipocytes (P<0.01). The sequence of goat KLF6 gene was obtained and its molecular characteristics were identified. It was found that it was highly expressed in adipose tissue, and the expression level after differentiation was significantly higher than that before differentiation.It provides important data for revealing the regulation of KLF6 on the differentiation of goat adipocytes.

Key words: Goat, KLF6, Tissue expression, Preadipocytes, Cell differentiation

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引用本文

王江林, 王永, 孟庆勇, 朱江江, 林亚秋. 山羊KLF6的分子克隆及其时空表达特征分析[J]. 华北农学报, 2021, 36(2): 226-232. doi: 10.7668/hbnxb.20191543.

WANG Jianglin, WANG Yong, MENG Qingyong, ZHU Jiangjiang, LIN Yaqiu. Molecular Cloning and Spatio-temporal Expression Analysis of KLF6 in Goat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(2): 226-232. doi: 10.7668/hbnxb.20191543.

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