牛miR-144靶基因预测与组织表达分析

doi:10.3969/j.issn.1004-1524.2022.03.07

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (3): 471-479.DOI: 10.3969/j.issn.1004-1524.2022.03.07

牛miR-144靶基因预测与组织表达分析

丁燕玲(), 王鹏飞, 杨朝云, 周小南, 赵志艳, 张岩峰, 史远刚, 康晓龙()

宁夏大学农学院,宁夏银川 750021

收稿日期:2021-06-08 出版日期:2022-03-25 发布日期:2022-03-30
通讯作者: 康晓龙
作者简介:*康晓龙,E-mail: kangxl9527@126.com
丁燕玲(1996—),女,甘肃兰州人,硕士研究生,研究方向为动物遗传育种与繁殖。E-mail: 1987560382@qq.com
基金资助:
国家自然科学基金(31760647);国家自然科学基金(31660645);宁夏自然科学基金(2020AAC03075);宁夏自然科学基金(2021AAC03027)

Prediction of target genes and tissue expression analysis of miR-144 in cattle

DING Yanling(), WANG Pengfei, YANG Chaoyun, ZHOU Xiaonan, ZHAO Zhiyan, ZHANG Yanfeng, SHI Yuan- gang, KANG Xiaolong()

School of Agriculture,Ningxia University, Yinchuan 750021, China

Received:2021-06-08 Online:2022-03-25 Published:2022-03-30
Contact: KANG Xiaolong

摘要/Abstract

摘要：

为研究miR-144的靶基因与miR-144在牛不同组织中的表达规律,预测其对肌肉生长发育的调节机制,对miR-144在各物种间的保守性、潜在靶基因及其富集通路进行分析,通过qRT-PCR方法检测牛不同组织中miR-144的表达。结果表明,miR-144成熟序列在各物种间保守性较高;富集分析发现,靶基因显著富集于血管平滑肌收缩、cGMP-PKG、cAMP等与肌肉发育相关通路中。qRT-PCR结果显示,miR-144在肝中的表达量最高,在皮下脂肪中的表达量最低;miR-144在腿肌和心中的表达差异不显著(P>0.05),但在肝中的表达显著(P<0.05)高于腿肌和心;miR-144在背最长肌中的表达显著(P<0.05)高于瘤胃和皮下脂肪,而瘤胃和皮下脂肪组织间表达差异不显著(P>0.05)。

关键词: miR-144, 组织表达, 靶基因预测, 实时荧光定量PCR, 富集通路

Abstract:

To study the target genes of miR-144 and their expression patterns in different tissues of cattle, and to predict the regulatory mechanism of miR-144 on muscle growth and development, the conservation, potential target genes and their enrichment pathways of miR-144 among different species were analyzed, and expression of miR-144 in different tissues of cattle was detected by qRT-PCR. The results showed that mature sequence of miR-144 was highly conserved among species; Enrichment analysis showed that target genes were significantly enriched in vascular smooth muscle contraction, cGMP-PKG, cAMP and other pathways related to muscle development. qRT-PCR results showed that miR-144 had the highest expression in liver and the lowest expression in subcutaneous fat; There was no significant difference(P>0.05) in the expression of miR-144 in leg muscle and heart, but the expression of miR-144 in liver was significantly (P<0.05) higher than that in leg muscle and heart; The expression of miR-144 in longissimus dorsi was significantly (P<0.05) higher than that in rumen and subcutaneous adipose, but there was no significant difference (P>0.05) between rumen and subcutaneous adipose.

Key words: miR-144, tissue expression, target gene prediction, qRT-PCR, enrichment pathway

中图分类号:

S823

丁燕玲, 王鹏飞, 杨朝云, 周小南, 赵志艳, 张岩峰, 史远刚, 康晓龙. 牛miR-144靶基因预测与组织表达分析[J]. 浙江农业学报, 2022, 34(3): 471-479.

DING Yanling, WANG Pengfei, YANG Chaoyun, ZHOU Xiaonan, ZHAO Zhiyan, ZHANG Yanfeng, SHI Yuan- gang, KANG Xiaolong. Prediction of target genes and tissue expression analysis of miR-144 in cattle[J]. Acta Agriculturae Zhejiangensis, 2022, 34(3): 471-479.

图/表 8

参考文献 36

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数据库名称 Databasename	功能 Function	网址 Website
miRBase	miRNA序列数据库miRNA sequence database	http://www.mirbase.org/index.shtml/
TargetScan	miRNA靶基因预测Prediction of miRNA target genes	http://www.targetscan.org/
miRWalk	miRNA靶基因预测Prediction of miRNA target genes	http://mirwalk.umm.uni-heidelberg.de/
miRDB	miRNA靶基因预测Prediction of miRNA target genes	http://www.mirdb.org/
Veney	维恩图绘制Drawing of wenn diagram	http//bioinfogp.cnb.csic.es/tools/Venny/index.html
miRTarbase	miRNA实验验证靶基因数据库	http://mirtar-base.mbc.nctu.edu.tw/
	Validation of target genes database by miRNA experiment
DAVID	功能注释Function notes	https://david.ncifcrf.gov/

数据库名称 Databasename	功能 Function	网址 Website
miRBase	miRNA序列数据库miRNA sequence database	http://www.mirbase.org/index.shtml/
TargetScan	miRNA靶基因预测Prediction of miRNA target genes	http://www.targetscan.org/
miRWalk	miRNA靶基因预测Prediction of miRNA target genes	http://mirwalk.umm.uni-heidelberg.de/
miRDB	miRNA靶基因预测Prediction of miRNA target genes	http://www.mirdb.org/
Veney	维恩图绘制Drawing of wenn diagram	http//bioinfogp.cnb.csic.es/tools/Venny/index.html
miRTarbase	miRNA实验验证靶基因数据库	http://mirtar-base.mbc.nctu.edu.tw/
	Validation of target genes database by miRNA experiment
DAVID	功能注释Function notes	https://david.ncifcrf.gov/

基因 Gene	引物序列(5'→3') Primer sequence(5'→3')	引物长度 Primer length/bp
miR-144	RTprimer:GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTAGTA	50
	F:CGCGCGTACAGTATAGATGATG	23
	R:AGTGCAGGGTCCGAGGTATT	20
18S RNA	F:GTGGTGTTGAGGAAAGCAGACA	22
	R:TGATCACACGTTCCACCTCATC	22

基因 Gene	引物序列(5'→3') Primer sequence(5'→3')	引物长度 Primer length/bp
miR-144	RTprimer:GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTAGTA	50
	F:CGCGCGTACAGTATAGATGATG	23
	R:AGTGCAGGGTCCGAGGTATT	20
18S RNA	F:GTGGTGTTGAGGAAAGCAGACA	22
	R:TGATCACACGTTCCACCTCATC	22

物种Species	登录号Accession No.	序列Sequence(5'~3')
牛Bos taurus	MIMAT0009234	UACAGUAUAGAUGAUGUACUAG
马Equus caballus	MIMAT0013024	UACAGUAUAGAUGAUGUACU
猪Sus scrofa	MIMAT0025364	UACAGUAUAGAUGAUGUAC
狗Canis lupus familiaris	MIMAT0006734	UACAGUAUAGAUGAUGUACUAG
鸡Gallus	MIMAT0003776	CUACAGUAUAGAUGAUGUACUC
小鼠Mus musculus	MIMAT0000156	UACAGUAUAGAUGAUGUACU
家鼠Rattus norvegicus	MIMAT0000850	UACAGUAUAGAUGAUGUACU
人Homo sapiens	MIMAT0000436	UACAGUAUAGAUGAUGUACU
黑猩猩Pan troglodytes	MIMAT0002262	UACAGUAUAGAUGAUGUACUAG
斑马鱼Brachydaniorerio var	MIMAT0001841	UACAGUAUAGAUGAUGUACU

牛miR-144靶基因预测与组织表达分析

Prediction of target genes and tissue expression analysis of miR-144 in cattle

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