牛FOXP2基因的单等位基因表达和DNA甲基化状态分析

doi:10.11843/j.issn.0366-6964.2025.09.020

Abstract

Abstract:

The aim of this study was to investigate the monoallelic expressions and DNA methylation status of the forkhead box P2 (FOXP2) gene in bovine tissues and placentas. The bovine tissues of two growth periods (calves of 0 d old and adult cows of 4-5 years old) and full-term placentas naturally delivered were collected. The expression levels of the FOXP2 gene were analyzed in bovine 6 tissues (heart, liver, spleen, lung, kidney and brain) and placentas by qRT-PCR. The monoallelic expressions of the FOXP2 gene were analyzed using the method of direct sequencing of RT-PCR products based on the single nucleotide polymorphism (SNP). The analysis of DNA methylation status in bovine tissues, placentas and sperms was conducted using sulphite sequencing method. The results showed that the expression of FOXP2 gene in all 6 tissues (heart, liver, spleen, lung, kidney and brain) of adult bovine was higher than that of calf tissues. Based on the SNP site of g.53771685G > A situated in the 3' non-coding region of the FOXP2 gene, monoallelic expression of FOXP2 was observed in 6 tissues of calves. However, in adult cattle, FOXP2 showed monoallelic expression only in brain tissue and biallelic expression in other 5 tissues (heart, liver, spleen, lung and kidney). The FOXP2 gene was mathernally expressed in the placenta. Hypomethylation was showed at the promoter and the 5' end of FOXP2 X1 splice in bovine tissues, placentas and sperms. These results indicated that the FOXP2 gene is an imprinted gene and the maternal allele is expressed in bovine placenta, and the monoallelic expression in bovine tissues presents growth period- and tissue-specific. Hypomethylation is in the promoter and 5' regions of FOXP2 gene, indicating that DNA methylation is not involved in the regulating monoallelic expression of bovine FOXP2 gene.

Key words: forkhead box P2(FOXP2) gene, allelic expression, DNA methylation, bovine

CLC Number:

ZHENG Yunchang, HOU Ruilin, LIANG Xiaohe, YANG Lidan, ZHANG Yinjiao, HUO Haonan, CHEN Weina, ZHANG Cui, LI Shijie. Analysis of Monoallelic Expression and DNA Methylation Status of Bovine FOXP2 Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4369-4378.

Figures/Tables 5

Table 1

Fig. 1

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Fig. 4

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引物名称 Primer name	引物序列(5′→3′) Primer sequence	用途 Application	扩增片段/bp Amplicon size	退火温度/℃ Tm	PCR类型 Type of PCR
q-F	CAGTACGTCATAATCTTAGCCT	表达量的检测	316	60	cDNA
q-R	CCATTGCTGTCGATATGATCC
GA-F	AACCACGAGAAGTATAAC	内参基因	223	60	cDNA
GA-R	AAGCAGGGATGATATTC
g-F	CCAATCCTTCCCTCCTCAA	鉴定杂合子	659	52	DNA
g-R	TAATGAAACTAACACTATTTTTGG
c-F	GACAGCAATGGGAACAG	等位基因分析	619	53	cDNA
c-R	CATTTGCTTTCTACAGGATG
FO20-OF-1	GGTTGGGTTTGTATTGTGGA	甲基化分析	458	52
FO20-OR-1	TTTCACTCTATACACAAAAACAC
FO20-IF-1	GTTGGAAGGGAGGTTATAGT		372	53
FO20-IR-1	TCCTATAAATACAAACAACCAAAC
FOXP2-OF	GTGTGTGTTTGTGTATTAGT		595	52
FOXP2-OR	AAAAAAAAAAAACTACCAAAACTACC
FOXP2-IF	TTAAAGGTAGAAGTTGGTGTTGT		270	53
FOXP2-IR1	ACTACCAACCCCTCCCCTATC
FO249-OF-1	GGTGTGTTGTTTGTGGGTTT		478	56
FO249-OR-1	ACACATCTACTATTTCCACACACA
FO249-IF-1	GTAAGTTTGTTTATTTTTATTTTGG		293	53
FO249-IR-2	AAACAACCTCCCCTCTCTAA

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Analysis of Monoallelic Expression and DNA Methylation Status of Bovine FOXP2 Gene

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