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华贵栉孔扇贝MSTN基因启动子的功能分析

范嗣刚 赵超 王鹏飞 闫路路 邱丽华

范嗣刚, 赵超, 王鹏飞, 闫路路, 邱丽华. 华贵栉孔扇贝MSTN基因启动子的功能分析[J]. 南方水产科学, 2019, 15(1): 63-68. doi: 10.12131/20180182
引用本文: 范嗣刚, 赵超, 王鹏飞, 闫路路, 邱丽华. 华贵栉孔扇贝MSTN基因启动子的功能分析[J]. 南方水产科学, 2019, 15(1): 63-68. doi: 10.12131/20180182
Sigang FAN, Chao ZHAO, Pengfei WANG, Lulu YAN, Lihua QIU. Functional analysis of MSTN promoter in scallop (Chalmys nobilis)[J]. South China Fisheries Science, 2019, 15(1): 63-68. doi: 10.12131/20180182
Citation: Sigang FAN, Chao ZHAO, Pengfei WANG, Lulu YAN, Lihua QIU. Functional analysis of MSTN promoter in scallop (Chalmys nobilis)[J]. South China Fisheries Science, 2019, 15(1): 63-68. doi: 10.12131/20180182

华贵栉孔扇贝MSTN基因启动子的功能分析

doi: 10.12131/20180182
基金项目: 中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助 (2017YB10);广东省海洋渔业科技与产业发展专项 (A201401A07;B201601-Z01);海南省自然科学基金项目(20154180)
详细信息
    作者简介:

    范嗣刚(1982—),男,博士,助理研究员,从事海洋生物功能基因研究。E-mail: fansigang@scsfri.ac.cn

    通讯作者:

    邱丽华(1971 — ),女,博士,研究员,从事海洋生物功能基因研究。E-mail: qiugroup_bio@outlook.com

  • 中图分类号: S 917.4

Functional analysis of MSTN promoter in scallop (Chalmys nobilis)

  • 摘要: 为了解肌肉生长抑制素myostatin (MSTN)在华贵栉孔扇贝(Chalmys nobilis)闭壳肌肌肉生长和发育过程中所起的负调控作用,对华贵栉孔扇贝的MSTN启动子序列进行了生物信息学分析。结果显示,MSTN启动子序列长1 358 bp,有4个转录起始位点。核心启动子区为–100~–51 bp。有1个TATA-box (–92~–86 bp)和2个E-box等顺式作用元件;潜在的转录因子结合位点有MEF2、MEF3、FoxO、MTBF和MyoD等;启动子区域无CpG岛。成功构建了6个MSTN启动子不同长度片段的荧光素酶表达载体,瞬时转染到293T细胞并进行双荧光素酶报告基因活性检测,表明6个启动子片段均有转录活性,PGL-534的活性最高,其次为PGL-274、PGL-22和PGL-102,最低的为PGL-995。–216~–364区域可能存在负调控基因表达的转录因子结合位点,–364~–825区域可能存在正调控基因表达的转录因子结合位点。
  • 图  1  MSTN启动子序列分析

    下划直线表示转录因子结合位点,下划波浪线为顺式作用元件,斜体表示核心启动子区域,阴影部分表示转录起始位点

    Figure  1.  Analysis of MSTN gene promoter sequence

    The potential transcription factor binding sites are straightly underlined. The cis-regulatory elements are wavily underlined. Core promoter region is in italic. Nucletide of each transcription start sites are in shadow.

    图  2  MSTN基因启动子不同长度片段的扩增电泳图

    电泳顺序:PGL-22、PGL-102、PGL-274、PGL-534、PGL-995、PGL-1143、DL2 000

    Figure  2.  Electrophoretograms of MSTN gene promoter with different lengths

    Electrophoresis order: PGL-22, PGL-102, PGL-274, PGL-534, PGL-995, PGL-1143, DL2 000

    图  3  MSTN启动子不同长度片段的相对活性

    相同字母表示差异不显著,不同字母表示差异显著(P<0.05)

    Figure  3.  Relative luciferase activity of MSTN gene with different lengths

    The same letters indicate no significant difference, while different letters indicate significant difference (P<0.05).

    表  1  实验所用引物

    Table  1.   Primers used in this experiment

    引物
    primer
    序列 (5'−3')
    sequence
    PGL-22 CGGggtaccACACGGCGAAAAAATGGAGC
    PGL-102 CGGggtaccGGATGCCATAAATCAAAACCACAAC
    PGL-274 CGGggtaccAAAACGCCGCCAAACG
    PGL-534 CGGggtaccACTTCAGGCTGTATCGCAAAT
    PGL-995 CGGggtaccACCCGTTGGCAGCGTTCA
    PGL-1143 CGGggtaccTCTAAATGCTAACCCTTGTGCTG
    P1322A CTAaagcttTATAGCGGTTACGTTACAGATGGTT
     注:小写字体为酶切位点  Note: Lowercase letters represent restriction enzyme loci.
    下载: 导出CSV
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出版历程
  • 收稿日期:  2018-08-16
  • 修回日期:  2018-09-15
  • 网络出版日期:  2018-12-14
  • 刊出日期:  2019-02-05

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