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| Relationship Between the Methylation Level of VVD Candidate Gene ST3GAL4 and ALP Activity and Gene Expression in Chicken (Gallus gallus) |
| MA Yan-Chao1, LI Jian-Zeng1, ZHANG Lu-Jie1, CAI Chun-Xia1, JIANG Rui-Rui1, ZHANG Yan-Hua1, TIAN Ya-Dong1, HAN Lu2, JI Jin-Qing2, HAN Rui-Li1,* |
1 College of Animal Science and Technology/Key Laboratory of Livestock Resources (Poultry) Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Henan Agricultural University, Zhengzhou 450046, China;
2 Henan Animal Husbandry Station, Zhengzhou 450008, China |
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Abstract Valgus-varus Deformity (VVD) is one of the most common leg diseases in broiler (Gallus gallus) breeding industry. To explore the relationship between the methylation level of the promoter region of the VVD related gene ST3 β-galactoside α-2,3-sialyltransferase 4 (ST3GAL4) and the activity and gene expression of alkaline phosphatase (ALP), the differential expression of ST3GAL4 in different tissues was detected by qPCR, and the differential expression tissues were screened. Further, bisulfite-sequencing PCR (BSP) were carried out to detect the methylation level of ST3GAL4 in blood and differentially expressed tissues. The results showed that the ALP activity of broilers in VVD group was significantly lower than that in normal group (P<0.05); The expression level of ST3GAL4 gene in leg muscle of broilers in VVD group was significantly higher than that in normal group (P<0.05). The results of methylation level showed that the overall methylation level of blood ST3GAL4 promoter region in VVD group was higher than that in normal group (P=0.051), and the methylation level of CpG25 was significantly different (P<0.05). In leg muscle tissue, the overall methylation level of ST3GAL4 promoter region of broilers in VVD group was significantly lower than that in normal group (P<0.01), and the methylation levels of CpG11, CpG14 and CpG15 were significantly different (P<0.05). The prediction results of transcription factors in ST3GAL4 promoter region showed that the key site CpG15 might bind to the transcription factors cAMP response element binding protein 1 (CRE-BP1), cAMP response element binding protein (CREB) and cytoplasmic polyadenylation element binding protein (CPE bind), and the key site CpG25 might bind to the transcription factors specific protein 1 (Sp1) and activator protein-2α (AP-2α). Correlation analysis showed that the methylation level of CpG25 in the ST3GAL4 promoter region in blood was negatively correlated with serum ALP activity, and the methylation level of CpG15 in the ST3GAL4 promoter region in leg muscle was also negatively correlated with gene expression. This study reveals that the methylation level of the promoter region of ST3GAL4 gene in VVD broilers is correlated with gene expression level and serum ALP activity, which provides reference data for disease resistance breeding of broilers.
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Received: 25 April 2022
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Corresponding Authors:
*rlhan@126.com
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