miR-423-5p在牛肌肉组织中表达及其靶基因预测

doi:10.3969/j.issn.1004-1524.2021.05.02

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (5): 785-793.DOI: 10.3969/j.issn.1004-1524.2021.05.02

miR-423-5p在牛肌肉组织中表达及其靶基因预测

陈雯¹(), 张伟伟¹^,²^,^*(), 邵淑丽¹^,², 付学鹏¹^,², 黄鑫¹^,², 李铁¹^,²

1.齐齐哈尔大学生命科学与农林学院, 黑龙江齐齐哈尔 161006
2.抗性基因工程与寒地生物多样性保护黑龙江省重点实验室,黑龙江齐齐哈尔 161006

收稿日期:2020-11-13 出版日期:2021-05-25 发布日期:2021-05-25
通讯作者: 张伟伟
作者简介:*张伟伟,E-mail:zww121@163.com
陈雯(1995—),女,天津人,硕士研究生,研究方向为骨骼肌细胞增殖分化。E-mail:chenwen_1120@163.com
基金资助:
国家自然科学基金(31801148);黑龙江省自然科学基金(C2016057);黑龙江省教育厅基本业务专项(135209260);黑龙江省省属高等学校基本科研业务费科研项目(YSTSXK201874)

Expression of miR-423-5p in bovine muscle and predicted target genes

CHEN Wen¹(), ZHANG Weiwei¹^,²^,^*(), SHAO Shuli¹^,², FU Xuepeng¹^,², HUANG Xin¹^,², LI Tie¹^,²

1. College of Life Sciences, Agriculture and Forestry, Qiqihar University, Qiqihar 161006, China
2. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar 161006, China

Received:2020-11-13 Online:2021-05-25 Published:2021-05-25
Contact: ZHANG Weiwei

摘要/Abstract

摘要：

miR-423-5p在细胞中具有重要的生物学功能,如在肌细胞中能影响细胞的增殖。根据miRBase及NCBI网站显示的相关信息利用分析软件对miR-423在各物种之间的同源性进行了分析。取成年西门塔尔牛体内的骨骼肌、小肠、心脏组织利用茎环荧光定量PCR进行miR-423-5p表达量检测,将牛骨骼肌卫星细胞(MDSCs)分化处理不同时长后进行检测;最后通过在线软件(TargetScan、PicTar)预测其可能结合的靶基因。结果表明,miR-423-5p在各物种间具有较高的同源性,在骨骼肌中表达量明显高于其他组织。在MDSCs中,随着细胞分化时间的延长,miR-423-5p表达量逐渐升高。miR-423-5p在12个基因mRNA的3'-UTR上含有潜在的结合位点,其中部分靶基因参与细胞的增殖、分化和凋亡。结果提示,miR-423-5p可能参与细胞的生长代谢,调节细胞的增殖与分化。

关键词: miR-423-5p, 茎环荧光定量PCR, 靶基因预测, 牛

Abstract:

miR-423-5p has important biological functions in cells, for example, it can affect cell proliferation in myocytes. In this paper, homology analysis was performed based on the relevant information displayed on miRBase and NCBI websites using analysis software. Skeletal muscle, small intestine and heart tissues from adult Simmental cattle were taken for miR-423-5p expression assay using stem-loop real-time PCR, and the same assay was performed after differentiation of bovine skeletal muscle satellite cells (MDSCs) treated with different time. Finally, the possible binding target genes were predicted by online software (TargetScan, PicTar). The results showed that miR-423-5p had a high homology among species and significantly higher expressed in skeletal muscle than in other tissues. In MDSCs, miR-423-5p expression gradually increased as the differentiation time of cells increased. miR-423-5p contained potential binding sites on the 3'UTR of 12 gene mRNAs, some of the target genes were involved in cell proliferation, differentiation and apoptosis. The results suggested that miR-423-5p may be involved in cell growth metabolism and regulate cell proliferation and differentiation.

Key words: miR-423-5p, stem-loop real-time PCR, target gene prediction, bovine

中图分类号:

S823

陈雯, 张伟伟, 邵淑丽, 付学鹏, 黄鑫, 李铁. miR-423-5p在牛肌肉组织中表达及其靶基因预测[J]. 浙江农业学报, 2021, 33(5): 785-793.

CHEN Wen, ZHANG Weiwei, SHAO Shuli, FU Xuepeng, HUANG Xin, LI Tie. Expression of miR-423-5p in bovine muscle and predicted target genes[J]. Acta Agriculturae Zhejiangensis, 2021, 33(5): 785-793.

图/表 8

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引物名称 Primer name	引物序列 Primer sequence
bta-miR-423 RT Primer	5'-CTC AAC TGG TGT CGT GGA GTC GGC AAT TCA GTT GAG AAA GTC TC-3'
bta-miR-423-5p Forward primer	5'-ACA CTC CAG CTG GGT GAG GGG CAG AGA GCG AGA-3'
bta-miR-423-5p Reverse primer	5'-CTC AAC TGG TGT CGT GGA-3'
18S Forward primer	5'-GGA CAT CTA AGG GCA TCA CAG-3'
18S Reverse primer	5'-AAT TCC GAT AAC GAA CGA GAC T-3'

引物名称 Primer name	引物序列 Primer sequence
bta-miR-423 RT Primer	5'-CTC AAC TGG TGT CGT GGA GTC GGC AAT TCA GTT GAG AAA GTC TC-3'
bta-miR-423-5p Forward primer	5'-ACA CTC CAG CTG GGT GAG GGG CAG AGA GCG AGA-3'
bta-miR-423-5p Reverse primer	5'-CTC AAC TGG TGT CGT GGA-3'
18S Forward primer	5'-GGA CAT CTA AGG GCA TCA CAG-3'
18S Reverse primer	5'-AAT TCC GAT AAC GAA CGA GAC T-3'

序号 No.	名称 Name		前体序列 Precursor sequence			成熟序列 Mature sequence		基因组位置 Genomic location
1	hsa-mir- 423		AUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUAACCCGCGC			>hsa-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >hsa-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr17: 30117079-30117172 [+]
2	mmu-mir- 423		ACUUGUGAGGAAAUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGCUUG			>mmu-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >mmu-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr11: 77078064-77078172 [-]
3	bta-mir- 423		AUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGC			>bta-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >bta-miR-423-3p AAGCUCGGUCUGAGGCCCCUCAGU		chr19: 21923268-21923361 [+]
4	cgr-mir- 423		GAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCC			>cgr-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >cgr-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		JH000631.1: 520509-520586 [-]
5	ssc-mir- 423		AAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUAC			>ssc-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >ssc-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr12: 48287019-48287098 [-]
6	eca-mir- 423		AGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU			>eca-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >eca-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr11: 43782441-43782503 [+]
7	chi-mir- 423		ACUUGUGAGGAAAUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGCUUGAGUUUCUCCCC			>chi-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >chi-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr19: 21574266-21574385 [+]
序号 No.		名称 Name		前体序列 Precursor sequence	成熟序列 Mature sequence		基因组位置 Genomic location
8		pal-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>pal-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >pal-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		KB031150.1: 20857586-20857644 [+]
9		ocu-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>ocu-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >ocu-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		CM000808.1: 18146102-18146160 [-]
10		dno-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>dno-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >dno-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		JH582994.1: 2753245-2753303 [-]
11		rno-mir- 423		GAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCU	>rno-miR-423-5p UGAGGGGCAGAGAGCGAGACUUUU >rno-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr10: 63098366-63098444 [+]
12		cpo-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>cpo-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >cpo-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		DS562887.1: 22509141-22509199 [-]
13		mml-mir- 423		AUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGC	>mml-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >mml-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr16: 24183848-24183941 [+]
14		ppy-mir- 423		AGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGC	>ppy-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >ppy-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr17:4902242-24902331 [+]

序号 No.	名称 Name		前体序列 Precursor sequence			成熟序列 Mature sequence		基因组位置 Genomic location
1	hsa-mir- 423		AUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUAACCCGCGC			>hsa-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >hsa-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr17: 30117079-30117172 [+]
2	mmu-mir- 423		ACUUGUGAGGAAAUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGCUUG			>mmu-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >mmu-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr11: 77078064-77078172 [-]
3	bta-mir- 423		AUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGC			>bta-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >bta-miR-423-3p AAGCUCGGUCUGAGGCCCCUCAGU		chr19: 21923268-21923361 [+]
4	cgr-mir- 423		GAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCC			>cgr-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >cgr-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		JH000631.1: 520509-520586 [-]
5	ssc-mir- 423		AAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUAC			>ssc-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >ssc-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr12: 48287019-48287098 [-]
6	eca-mir- 423		AGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU			>eca-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >eca-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr11: 43782441-43782503 [+]
7	chi-mir- 423		ACUUGUGAGGAAAUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGCUUGAGUUUCUCCCC			>chi-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >chi-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr19: 21574266-21574385 [+]
序号 No.		名称 Name		前体序列 Precursor sequence	成熟序列 Mature sequence		基因组位置 Genomic location
8		pal-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>pal-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >pal-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		KB031150.1: 20857586-20857644 [+]
9		ocu-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>ocu-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >ocu-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		CM000808.1: 18146102-18146160 [-]
10		dno-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>dno-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >dno-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		JH582994.1: 2753245-2753303 [-]
11		rno-mir- 423		GAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCU	>rno-miR-423-5p UGAGGGGCAGAGAGCGAGACUUUU >rno-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr10: 63098366-63098444 [+]
12		cpo-mir- 423		UGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGU	>cpo-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >cpo-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		DS562887.1: 22509141-22509199 [-]
13		mml-mir- 423		AUAAAGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGC	>mml-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >mml-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr16: 24183848-24183941 [+]
14		ppy-mir- 423		AGGAAGUUAGGCUGAGGGGCAGAGAGCGAGACUUUUCUAUUUUCCAAAAGCUCGGUCUGAGGCCCCUCAGUCUUGCUUCCUACCCCGCGC	>ppy-miR-423-5p UGAGGGGCAGAGAGCGAGACUUU >ppy-miR-423-3p AGCUCGGUCUGAGGCCCCUCAGU		chr17:4902242-24902331 [+]

序号 No	靶基因 Target gene	靶基因名称 The name of target gene	与靶基因mRNA 3'UTR结合位点 The binding site on 3'UTRs of target genes mRNA
1	PEX26	过氧化物酶体生物发生因子26 Peroxisomal biogenesis factor 26	560~566
2	THBS3	血小板反应蛋白3 Thrombospondin 3	76~83
3	CLTB	网格蛋白轻链B Clathrin light chain B	153~159
4	OPA3	线粒体外膜脂质代谢调节剂OPA3	171~177
		Outer mitochondrial membrane lipid metabolism regulator OPA3
5	LYNX1	Ly6/神经毒素1 Ly6/neurotoxin 1	705~711
6	SRM	亚精胺合酶 Spermidine synthase	232~239
7	VGF	VGF可诱导神经生长因子 VGF nerve growth factor inducible	415~421
8	CAMKK1	钙/钙调蛋白依赖性蛋白激酶激酶1	61~67
		Calcium/calmodulin dependent protein kinase kinase 1
9	WNT9B	Wnt家庭成员9B Wnt family member 9B	222~228
10	PLA2G6	磷脂酶A2组Ⅵ Phospholipase A2 group Ⅵ	424~430
11	NRSN2	神经素2 Neurensin 2	843~849
12	SYP	突触素 Synaptophysin	1 329~1 335

miR-423-5p在牛肌肉组织中表达及其靶基因预测

Expression of miR-423-5p in bovine muscle and predicted target genes

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