山麻鸭SCA-2基因克隆、生物信息学及组织表达谱分析  

Cloning, bioinformatics and tissue expression profile analysis of SCA-2 gene in Shan partridge ducks

作　　者：张迎燕 刘亚楠 江林 梅慧灵 郑定黔 田若宁 徐丽 李昂[1] 吴旭[1] ZHANG Yingyan;LIU Yanan;JIANG Lin;MEI Huiling;ZHENG Dingqian;TIAN Ruoning;XU Li;LI Ang;WU Xu(College of Animal Science(College of Bee Science),Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福州350002

出　　处：《黑龙江畜牧兽医》2022年第6期47-52,135,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：福建省自然科学基金项目(2020J01536);福建省农业良种育种攻关与产业化工程项目(zycxny2021014);现代农业产业技术体系建设专项基金项目(CARS-43);国家自然科学基金项目(31001005)。

摘　　要：为了探究Struthiocalcin-2(SCA-2)基因在山麻鸭蛋壳形成过程中的作用,试验选择9只产蛋高峰期(30周龄)的健康山麻鸭,采用SMART-RACE方法对山麻鸭SCA-2基因cDNA序列进行克隆,运用生物信息学软件对SCA-2基因编码蛋白质的理化性质、二级和三级结构进行预测及分析,构建SCA-2基因系统进化树;采集试验山麻鸭心脏、肝脏、脾脏、肺脏、肾脏、胸肌、肌胃、下丘脑、垂体、卵巢,以及输卵管的漏斗部、膨大部、峡部和子宫部共14个组织,每3只做一次混样处理,每个组织共3个重复,提取组织RNA,通过实时荧光定量PCR分析SCA-2基因mRNA在上述组织中的相对表达量。结果表明:SCA-2基因的cDNA全长为726 bp,该基因编码蛋白的分子式为C_(836)H_(1247)N_(229)O_(247)S_(8),属于不稳定蛋白,其中丝氨酸(12.6%)、亮氨酸(12.0%)、丙氨酸(10.2%)含量较高;蛋白质的二级结构中,α螺旋占23.35%、β折叠占13.77%、无规卷曲占62.87%,且包含信号肽序列,不含跨膜结构;山麻鸭与绿头鸭、凤头潜鸭和棕硬尾鸭的亲缘关系最近;SCA-2基因在山麻鸭14种组织中均有不同程度的表达,其中在输卵管子宫部和峡部组织中的表达量极显著高于卵巢(P<0.01),明显高于其他组织。说明本试验成功克隆到了SCA-2基因,该基因可能在山麻鸭蛋壳的形成过程中扮演着重要角色。In order to explore the role of Struthiocalcin-2(SCA-2) gene in the formation of the eggshell of Shan partridge ducks, in the experiment, 9 healthy Shan partridge ducks during the peak period of egg production(30 weeks of age) were selected, and the SMART-RACE method was used to clone and obtain the full-length sequence of SCA-2 gene cDNA of Shan partridge duck.Bioinformatics software was used to predict and analyze the physical and chemical properties of the amino acids encoded by the SCA-2 gene and the secondary and tertiary structure of the protein, the phylogenetic tree of the SCA-2 gene was carried out. Fourteen tissues including heart, liver, spleen, lung, kidney, pectoralis, gizzard, hypothalamus, pituitary, ovary, and infundibular, bulge, isthmus and uterus of oviduct were collected.Every 3 samples were mixed once, with 3 replicate groups in total.These samples were used for RNA tissue extraction. The relative expression of SCA-2 gene in different tissues was analyzed. The results showed that the full length of the cDNA of the SCA-2 gene was 726 bp.The molecular formula of the protein encoded by the gene was C_(836)H_(1247)N_(229)O_(247)S_(8), which was an unstable protein with high contents of serine(12.6%), leucine(12.0%), and alanine(10.2%). In the secondary structure of the protein, α helix accounted for 23.35%, β sheet accounted for 13.77%, and random coil accounted for 62.87%, and contained peptide signal, but did not contain transmembrane structure.There are the closest genetic relationship between the Shan partridge duck and the Anas platyrhynchos, Aythyafuligula, and Oxyura jamaicensis. Different levels of SCA-2 gene expression were found in 14 tissues of the Shan partridge duck, and the expression level in the uterus and isthmus of oviduct was significantly higher than that in ovary(P<0.01). It indicated that the SCA-2 gene was successfully cloned in this experiment, and the gene may play an important role in the formation of the eggshell of Shan partridge ducks.

关 键 词：山麻鸭 SCA-2基因 基因克隆 生物信息学 组织表达谱 

分 类 号：S834.2[农业科学—畜牧学]