摘要
旨在克隆山羊SAMD12基因,明确SAMD12分子特征及表达特征,探究SAMD12基因对山羊肌内脂肪细胞分化的影响.首先通过RT-PCR技术扩增获得山羊SAMD12基因序列,利用生物信息学分析软件及qPCR技术明确山羊SAMD12分子特征及其在山羊组织、肌内脂肪细胞分化过程中的表达模式.结果表明,扩增获得山羊SAMD12基因序列734 bp, CDS区486 bp,编码氨基酸161个.山羊SAMD12特征分析结果显示SAMD12蛋白带正电且属于碱性亲水不稳定蛋白,主要分布于细胞核(52.2%),含有SAM家族所特有的SAM功能结构域.SAMD12蛋白二级结构中大部分氨基酸以α-螺旋、无规卷曲的方式存在.同源性比对结果显示山羊SAMD12与绵羊SAMD12同源性高达90.7%,同源性最低的为马(23.4%).系统进化树结果显示:山羊与绵羊、家牛划分为一支,说明山羊SAMD12与这两种哺乳动物亲缘关系最近.组织表达特征分析显示:SAMD12在山羊多个组织中均有表达,在瘤胃中表达量最高,其次为肾,这两种组织中的表达量都显著高于其他组织(P<0.05).细胞时序表达特征谱分析显示:成脂诱导分化后山羊SAMD12基因整体表达水平呈上升趋势,在诱导分化96 h时表达量达到最高值,显著高于0 h(P<0.05).以上结果提示,SAMD12基因在山羊肌内脂肪细胞成脂过程中可能起重要作用.
The purpose of this study was to clone goatSAMD12 gene,identify its molecular characteristics and expression char⁃acteristics,and explore the effect ofSAMD12 gene on the differentiation of goat intramuscular adipocytes.The goatSAMD12 gene sequence was amplified by RT⁃PCR.The molecular characteristics of goatSAMD12 and its expression pattern in the differentia⁃tion process of goat tissues and intramuscular adipocytes were identified by bioinformatics analysis software and qPCR technolo⁃gy.The results showed that the goatSAMD12 gene sequence was 734 bp,the CDS region was 486 bp,encoding 161 amino acids.The characteristic analysis of goatSAMD12 showed that SAMD12 protein was positively charged and belonged to basic hy⁃drophilic unstable protein,mainly distributed in the nucleus(52.2%)and contained SAM functional domain specific to SAM family.The prediction results of the secondary structure of SAMD12 protein showed that most amino acids existed in alpha helix and random coil.The results of homology comparison showed that goatSAMD12 had 90.7%homology with sheepSAMD12,and horseSAMD12(23.4%)had the lowest homology.The phylogenetic tree showed that goat,sheep and cattle were divided into one branch,indicating that goat SAMD12 had the closest genetic relationship with these two mammals.The analysis of tissue ex⁃pression characteristics showed thatSAMD12 was expressed in many tissues,with the highest expression in the rumen,followed by the kidney.The expression ofSAMD12 in these two tissues was significantly higher than that in other tissues(P<0.05).Temporal expression spectrum analysis showed that the overall expression level ofSAMD12 gene in cells after adipogenic differ⁃entiation showed an upward trend,and the expression reached the highest value at 96 h of differentiation induction,significantly higher than that at 0 h(P<0.05).The above results suggest thatSAMD12 may play a role in promoting intramuscular adipo⁃genesis in goats.
作者
吕锦诗
李安
刘珂含
王永
李艳艳
王友利
刘伟
林亚秋
LV Jin-shi;LI An;LIU Ke-han;WANG Yong;LI Yan-yan;WANG You-li;LIU Wei;LIN Ya-qiu(Ministry of Education Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Chengdu 610041,China;Sichuan Province Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Chengdu 610041,China;School of Animal Science and Veterinary Medicine,Southwest Minzu University,Chengdu 610041,China)
出处
《西南民族大学学报（自然科学版）》
CAS
2023年第2期134-141,共8页
Journal of Southwest Minzu University(Natural Science Edition)
基金
四川省青年科技创新团队:肉羊种质资源创新与利用四川省青年科技创新研究团队(2022JDTD0030)
西南民族大学研究生创新型科研项目(CX2021SZ68)。
关键词
山羊
SAMD12基因
生物学特性
表达特征
goat
SAMD12 gene
biological characteristics
expression characteristics
