| 摘要: |
| 【目的】克隆牦牛BTG2基因编码区（CDS区），对其进行生物信息学分析和组织表达分析，为后续深入研究牦牛BTG2基因的生物学功能奠定基础。【方法】以美仁牦牛为研究对象，采集其心脏、肝脏、脾脏、肌肉、脂肪和睾丸组织。提取脂肪组织总RNA，反转录获得cDNA。PCR克隆牦牛BTG2基因的CDS 区，测序后进行生物信息学分析。利用RT-qPCR技术，分析BTG2基因在心脏、肝脏、脾脏、肌肉、脂肪和睾丸组织中的表达水平。【结果】牦牛BTG2基因的CDS区全长453 bp，共编码150个氨基酸；BTG2蛋白的分子式为C748H1190N208O213S8，分子质量为16 761.42 ku，原子总数为2 367个；不稳定性指数为48.51，是不稳定蛋白；理论等电点为9.14；主要分布于细胞核（占56.5%）和线粒体（占26.1%）；氨基酸序列的1～108位有1个BTG1域；总平均亲水系数为－0.123，是亲水蛋白；共有15个磷酸化位点；二级结构以α-螺旋和无规则卷曲为主；无跨膜结构且无信号肽区域；与10个蛋白存在互作关系。系统进化分析结果显示，美仁牦牛与野牦牛亲缘关系最近，与间蜂猴亲缘关系最远。RT-qPCR结果显示，BTG2在成年公牦牛的心脏、肝脏、脾脏、肌肉、脂肪和睾丸组织中均有表达，其中在脂肪组织和脾脏组织中表达水平较高，在肌肉和睾丸组织中表达水平较低。【结论】克隆了牦牛BTG2基因，对其进行了生物信息学分析，明确了其组织表达情况。 |
| 关键词: 美仁牦牛 BTG2基因 生物信息学分析 组织表达分析 系统进化 |
| DOI：DOI:10.13207/j.cnki.jnwafu.2025.06.003 |
| 分类号: |
| 基金项目:合作市牦牛种质提升与提质增效项目;甘肃省民生科技专项（21CX6NP219）；现代肉牛牦牛产业技术体系项目（CARS-37）；中国农业科学院创新工程项目（25-LIHPS-01） |
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| Cloning，bioinformatics，and tissue expression analysis of yak BTG2 gene |
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SHEN Jinwei1,2,3, ZHAO Xue4, LU Jianwei5, ZHA Lao5, YANG Shumeng6, LIANG Chunnian1,2, CHENG Shuru1
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1.College of Animal Science and Technology,Gansu Agricultural University，Lanzhou，Gansu 730070，China;2.Key Laboratory of Yak Breeding Engineering of Gansu Province,Lanzhou Institute of Animal Husbandry and Veterinary Medicine,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu 730050,China;3.Key Laboratory of Livestock and Poultry Genetics and Breeding on the Qinghai-Tibetan Plateau,Ministry of Agriculture and Rural Development,Lanzhou,Gansu 730050,China;4.Agricultural and Livestock Product Quality and Safety Testing and Inspection Center for Gannan Tibetan Autonomous Prefecture,Hezuo,Gansu 747000,China;5.Animal Husbandry Station,Zuogaiduoma Township for Gannan Tibetan Autonomous Prefecture,Hezuo,Gansu 747000,China;6.Animal Husbandry Workstation for Gannan Tibetan Autonomous Prefecture,Hezuo,Gansu 747000,China
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| Abstract: |
| 【Objective】This research aimed to clone the coding region (CDS region) of yak BTG2 gene,to analyze its bioinformatics and tissue expression,and to provide theoretical basis for the subsequent in-depth study of the biological function of yak BTG2 gene.【Method】Heart,liver,spleen,muscle,fat and testis tissues were collected from Meiren yaks.The total RNA of adipose tissue was extracted,and cDNA was obtained by reverse transcription.The CDS region of yak BTG2 gene was cloned by PCR,and then sequenced,and bioinformatic analysis was conducted.The expression levels of BTG2 gene in heart,liver,spleen,muscle,adipose and testis tissues were analyzed by RT-qPCR.【Result】The CDS region of yak BTG2 gene was 453 bp in length,encoding a total of 150 amino acids.The molecular formula of BTG2 protein was C748H1190N208O213S8,the molecular weight was 16 761.42 ku,and the total number of atoms was 2 367.The instability index was 48.51,which made it an unstable protein.The theoretical isoelectric point was 9.14,mainly distributed in the nucleus (56.5%) and mitochondria (26.1%),with a BTG1 domain at positions 1-108 of the amino acid sequence.It was a hydrophilic protein with a total average hydrophilicity coefficient of －0.123.A total of 15 phosphorylation sites were observed.The secondary structure was dominated by α-helices and random curls.No transmembrane structure and signal peptide region was observed.Interaction with 10 proteins was observed.The results of phylogenetic tree showed that Meiren yak was most closely related to wild yak and least closely related to mesquite monkey.RT-qPCR results showed that protein was expressed in heart,liver,spleen,muscle,adipose and testis tissues of adult male bulls,with higher expression levels in adipose and spleen tissues,and lower expression levels in muscle and testis tissues.【Conclusion】The yak BTG2 gene was cloned, and bioinformatic analysis was performed to clarify its tissue expression. |
| Key words: Meiren yak BTG2 gene bioinformatics analysis tissue expression analysis systems evolution |
