Cloning, Expression Analysis and Functional Identification of SOCS3 Gene in Anhui White Goat (Capra hircus)
CUI Heng-Yuan1, MA Cun-Xia1, CUI Shuang-Shuang1, GUO Yu-Zhu1, LIU Hong-Yu1,2, ZHANG Yun-Hai1,2, SONG Ning1,2,*
1 College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China; 2 Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-breeding, Hefei 230036, China
Abstract Suppressor of cytokine signaling 3 (SOCS3) is a protein that responds to signal transduction and regulates milk protein synthesis. αS1-casein is one of the most important allergens in ruminant milk. However, the effect of SOCS3 on αS1-casein synthesis in goats (Capra hircus) is still unclear. To elucidate the gene sequence, structure and expression of SOCS3, and the effect of SOCS3 on αS1-casein expression in goat mammary gland, in this study, Anhui white goat mammary tissue was used as experimental material, SOCS3 gene coding region sequence was cloned by PCR. Bioinformatics analysis was conducted by online software, the expression of SOCS3 in different lactation periods was detected, interference and overexpression of SOCS3 were performed for functional analysis. The results showed that the complete coding region of SOCS3 gene in goats was 690 bp, encoding 229 amino acids. The molecular weight of SOCS3 protein was 25.09 kD, the theoretical isoelectric point was 8.97. There were 34 phosphorylation sites, no signal peptide and transmembrane helix structure, and SOCS3 was an unstable protein carrying positive charge. SOCS3 protein interacted with SOCS2, Janus kinase (JAK), tyrosine kinase 2 (TYK2), insulin receptor substrate 2 (IRS2), leptin receptor (LEPR) and interleukin 6 (IL6) and other proteins. SOCS3 gene of goat was most closely related to sheep, followed by cattle (Bos taurus). The expression of SOCS3 gene was the highest in the early and peak lactation periods. In goat mammary epithelial cells, interference with SOCS3 gene significantly up-regulated αS1-casein expression, and overexpression of SOCS3 gene significantly down-regulated αS1-casein expression. These results indicated that SOCS3 had a negative regulatory effect on αS1-casein synthesis in goats. This study provides a basis for studying the regulatory mechanism of SOCS3 gene in goat αS1-casein synthesis.
CUI Heng-Yuan,MA Cun-Xia,CUI Shuang-Shuang, et al. Cloning, Expression Analysis and Functional Identification of SOCS3 Gene in Anhui White Goat (Capra hircus)[J]. 农业生物技术学报, 2024, 32(9): 2071-2080.
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