山羊C/EBPα、C/EBPβ和C/EBPδ基因克隆及表达模式研究  

Study on Cloning and Expression Patterns of Goat C/EBPα,C/EBPβand C/EBPδGenes

作　　者：池永东[1,2] 王永 赵越[1,2] 朱江江 林亚秋[1,2] CHI Yongdong;WANG Yong;ZHAO Yue;ZHU Jiangjiang;LIN Yaqiu(School of Life Science and Technology,Southwest Minzu University, Chengdu 610041, China;Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Ministry of Education,Sichuan Province, Chengdu 610041, China)

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041 [2]青藏高原动物遗传资源保护与利用教育部,四川省重点实验室,四川成都610041

出　　处：《华北农学报》2020年第5期231-238,共8页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31672395);四川省应用基础研究计划重点项目(2018JY0036);西南民族大学中央高校基本科研业务费专项资金(2020PTJS15004)。

摘　　要：在克隆获得山羊C/EBPα、C/EBPβ和C/EBPδ基因序列基础上分析该家族成员序列特征,明确组织表达特性,并揭示3个基因在山羊肌内前体脂肪细胞分化过程中的表达情况。采用RT-PCR方法克隆山羊C/EBPα、C/EBPβ和C/EBPδ基因序列,利用相关的生物软件进行生物信息学分析,利用qPCR检测3个基因在山羊各组织及山羊肌内前体脂肪细胞成脂分化中的表达情况。克隆得到C/EBPα、C/EBPβ和C/EBPδ基因序列分别为1062,1056,873 bp,所编码的蛋白均为不稳定亲水碱性蛋白,均无信号肽和跨膜结构域。C/EBPα在皮下脂肪中表达水平最高,C/EBPβ和C/EBPδ均在肺中表达水平最高。在前体脂肪细胞分化过程中C/EBPα和C/EBPδ均在前体脂肪细胞分化前表达水平最高,随着分化时间的增加表达水平降低,C/EBPβ在肌内前体脂肪细胞中的表达随着诱导分化天数的增加而增加,在分化7 d时表达水平最高。基于以上研究,推测3个基因在山羊肌内脂肪细胞分化过程中发挥着重要作用。This experiment was conducted to study the sequence characteristics,physicochemical properties of goat C/EBPα,C/EBPβand C/EBPδgenes and proteins,and the expression patterns in goat tissues and in the differentiation of goat intramuscular precursor adipocytes.The C/EBPα,C/EBPβand C/EBPδof goat were cloned by RT-PCR method and the biological characteristics were analyzed using the relevant biological software.qPCR was used to detect the expression levels of C/EBPα,C/EBPβand C/EBPδmRNA in different tissues and preadipocyte of goat.The results showed that the sequence of C/EBPαwas 1062 bp,C/EBPβwas 1056 bp and C/EBPδwas 873 bp.The proteins encoded by C/EBPα,C/EBPβand C/EBPδwere all unstable hydrophilic basic proteins,and had no signal peptide and transmembrane domain.The expression of three genes was detected in different tissues of goats.The expression level of C/EBPαwas highest in subcutaneous fat,and the expression pattern of C/EBPβand C/EBPδwas similar,and the expression level was highest in lung.The expression patterns of C/EBPαand C/EBPδwere similar in the differentiation of adipose precursor cells,and the expression level was the highest at 0 d after differentiation,and the expression level was decreased with the increase of differentiation time.C/EBPβwas found in intramuscular precursor fat.The expression of C/EBPβin the cells increased with the number of days of induced differentiation,and the expression level was highest at 7 days of differentiation.Based on the above studies,it is speculated that the three genes play an important role in the process of goat adipose differentiation.

关 键 词：山羊 基因克隆 表达 细胞分化 

分 类 号：Q78[生物学—分子生物学] S826[农业科学—畜牧学]