研究简报 SHORT COMMUNICATION

通过测序与预测分析靶向奶牛乳腺κ-酪蛋白的miRNA

  • 李文清 ,
  • 王潇阳 ,
  • 李晨婉 ,
  • 张国只 ,
  • 赵亚宁 ,
  • 张宏禄 ,
  • 李万利
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  • 1. 河南农业大学生命科学学院, 郑州 450002;
    2. 河南省农业科学院畜牧兽医研究所, 郑州 450002
李文清(1979-),女,河南潢川人,副教授,博士,主要从事奶牛营养与牛奶品质研究。E-mail:liwenqing605@163.com

收稿日期: 2021-05-08

  网络出版日期: 2021-12-16

基金资助

国家自然科学基金青年科学基金项目(31501978);河南省高等学校重点科研项目(16A230010)

miRNA Targeting κ-Casein in Cow Mammary Gland Analyzed by Sequencing and Prediction

  • LI Wenqing ,
  • WANG Xiaoyang ,
  • LI Chenwan ,
  • ZHANG Guozhi ,
  • ZHAO Yaning ,
  • ZHANG Honglu ,
  • LI Wanli
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  • 1. College of Life Science, Henan Agricultural University, Zhengzhou 450002, China;
    2. Animal Husbandry and Veterinary Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China

Received date: 2021-05-08

  Online published: 2021-12-16

摘要

本研究旨在验证通过测序与预测分析靶向奶牛乳腺κ-酪蛋白(CSN3)的miRNA。应用乳腺组织的小RNA测序、TargetScan预测和RNAhybrid分析,筛选了4个可能靶向CSN3的miRNA,分别是bta-miR-33a、bta-miR-2284ab、bta-miR-193b和bta-miR-193a-3p。接着,通过荧光素酶报告试验检测CSN3-3'非编码区(3'UTR)和4种miRNA的结合情况,结果表明只有bta-miR-193b和bta-miR-193a-3p能与CSN3-3'UTR结合(P<0.01)。牛奶体细胞相关基因表达趋势进一步验证了bta-miR-193b和bta-miR-193a-3p与CSN3的靶向关系。这些结果提示,bta-miR-193b和bta-miR-193a-3p可直接靶向CSN3,这可以解释从泌乳高峰期至泌乳后期牛奶中CSN3蛋白总量的减少。本试验结果为乳蛋白的调控研究提供了依据,并为miRNA的功能研究铺垫了道路。

本文引用格式

李文清 , 王潇阳 , 李晨婉 , 张国只 , 赵亚宁 , 张宏禄 , 李万利 . 通过测序与预测分析靶向奶牛乳腺κ-酪蛋白的miRNA[J]. 动物营养学报, 2021 , 33(12) : 7131 -7139 . DOI: 10.3969/j.issn.1006-267x.2021.12.052

Abstract

The aim of this study was to identify the miRNA targeting κ-casein (CSN3) in cow mammary gland analyzed by sequencing and prediction. Using small RNA sequencing, TargetScan prediction and RNAhybrid analysis, four possible targeting miRNA were screened, namely bta-miR-33a, bta-miR-2284ab, bta-miR-193b and bta-miR-193a-3p. Luciferase reporter assay of the CSN3-3'untranslated region (3'UTR) and four kinds of miRNA showed that bta-miR-193b and bta-miR-193a-3p could bind to the CSN3-3'UTR (P<0.01). Expressional trend of relative gene from milk somatic cell further verified the targeting relationship among bta-miR-193b, bta-miR-193a-3p and CSN3. These findings suggeste that bta-miR-193b and bta-miR-193a-3p can directly target CSN3. This may explain the decrease in the total amount of CSN3 protein in milk from peak lactation to late lactation. Our results provide an evidence for the regulation of milk proteins and pave the way for miRNA function studies.

参考文献

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