夏南牛SOX6基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2023.07.006

Abstract

Abstract: 【Objective】 This study was aimed to clone the CDS sequence of RY-box transcription factor 6 (SOX6) gene in Xiannan cattle and carry out bioinformatics analysis, and explore the expression rules of SOX6 in different tissues of Xiannan cattle at different months and the differentiation of bovine primary skeletal muscle cells.【Method】 The CDS sequence of SOX6 gene was amplified by PCR using the cDNA template of leg muscle in Xianan cattle. Sequence similarity alignment and phylogentic tree contruction were carried out with other species, and the bioinformatics analysis of SOX6 protein was performed by the online software. The relative expression of SOX6 gene in different tissues of calves (0 month old), young cattle (12 months old) and adult cattle (24 months old) and different differentiation stages of bovine primary skeletal muscle cells were analyzed by Real-time quantitative PCR.【Result】 The CDS region of SOX6 gene in Xianan cattle was 2 526 bp in length, which encoded 841 amino acids.The results of phylogenetic tree showed that Xianan cattle had the closest relationship with yak and the farthest relationship with chicken.SOX6 protein was a weakly alkaline and hydrophilic protein with a molecular weight of 93.35 ku, which located in the nucleus without transmembrane structure and signal peptide.The secondary structure of SOX6 protein was mainly composed of alpha-helix, random coil, extended chain and beta-turn, accounting for 41.26%, 45.54%, 8.09% and 5.11%, respectively.Protein interaction analysis showed that SOX6 had interaction with type Ⅱ collagen α1 (COL2A1), chondroproteoglycan (ACAN), carboxy-terminal-binding protein 2 (CTBP2), and forkhead box 3(FOXJ3).The relative expression of SOX6 gene in leg muscle of Xianan cattle was the highest, and the expression in lung and spleen was the lowest.The expression level of leg muscle in 12 months old was significantly higher than that in 0 and 24 months old cattle (P<0.05).The expression of SOX6 gene increased gradually with the differentiation of bovine primary skeletal muscle cells, and the expression level of SOX6 gene on days 6 was significantly higher than that on days 0, 2, 4 and 8 (P<0.05).【Conclusion】 The CDS sequence of SOX6 gene in Xiannan cattle was successfully cloned, and its encoded protein was unstable, weakly alkaline and hydrophilic.The expression of SOX6 gene was significantly higher in leg muscle of different months than other tissues, and the expression in bovine primary skeletal muscle cells on days 6 of differentiation was significantly higher than that at other periods.The results provided a theoretical basis for further study on the regulatory role of SOX6 gene in the muscle development of Xiannan cattle.

Key words: Xianan cattle; SOX6 gene; cloning; bioinformatics; tissue expression

CLC Number:

XING Shanshan, LI Xin, YU Lei, XU Huifen, CAI Hanfang, LI Ming. Cloning, Bioinformatics Analysis and Tissue Expression of SOX6 Gene in Xianan Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2651-2660.

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Cloning, Bioinformatics Analysis and Tissue Expression of SOX6 Gene in Xianan Cattle

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