大恒799肉鸡Myoz1基因CDs区克隆、序列分析及其组织表达研究  

CDs Cloning,Sequence Analysis and Tissue Expression of Myoz1 Gene in Daheng 799 Broiler

作　　者：朱师良 余春林[1] 邱莫寒[1] 张增荣[1] 熊霞[1] 胡陈明[1,2] 杨礼 彭涵[1,2] 陈家磊 宋小燕[1] 夏波 刘思洋 蒋小松[1] 杜华锐[1,2] 李晴云 杨朝武[1] ZHU Shiliang;YU Chunlin;QIU Mohan;ZHANG Zengrong;XIONG Xia;HU Chenming;YANG Li;PENG Han;CHEN Jialei;SONG Xiaoyan;XIA Bo;LIU Siyang;JIANG Xiaosong;DU Huarui;LI Qingyun;YANG Chaowu(Key Laboratory of Animal Genetics and Breeding,Sichuan Animal Science Academy,Chengdu,Sichuan 610066;Sichuan Daheng Poultry Breeding Co.,Ltd.,Chengdu,Sichuan 610066)

机构地区：[1]四川省畜牧科学研究院动物遗传育种重点实验室,四川成都610066 [2]四川大恒家禽育种有限公司,四川成都610066

出　　处：《中国家禽》2024年第9期26-33,共8页China Poultry

基　　金：国家重点研发计划项目(2022YFD1601608);现代农业产业技术体系建设专项资金(CARS-41-G07);四川省区域创新合作项目(2023YFQ0035)。

摘　　要：为研究钙调神经磷酸酶肌小结结合蛋白1(Myoz1)的氨基酸序列特征及其在不同组织器官中的mRNA表达水平,试验通过PCR克隆Myoz1 CDs序列,分别利用ExPASYProt⁃param、ProtScale、Target P、DNAStar Protean、SWISS-MODEL、SignalP 4.1 Server、NetPhos2.0Serv⁃er和STRTNG软件,进行Myoz1蛋白理化特性、亲/疏水性、亚细胞结构、蛋白二级结构、三级结构、信号肽、磷酸化位点和蛋白互作分析,并通过实时荧光定量PCR(qRT-PCR)检测Myoz1基因表达量。结果显示:大恒799肉鸡Myoz1基因CDs序列全长为881 bp,可编码293个氨基酸;大恒799肉鸡Myoz1基因氨基酸序列与雉鸡、岩雷鸟位于一个分支,同源性最高,均高于96%,亲缘关系最近;Myoz1蛋白为水溶性蛋白且大多分布于细胞核内,分子式为C1442H_(2)244N400O430S10,分子质量约为32384.70 ku,理论等电点为9.12;Myoz1蛋白不存在信号肽,为分泌蛋白,共存在37个潜在的磷酸化位点、1个N-糖基化潜在位点;Myoz1蛋白二级结构主要由α-螺旋、β-转角和无规则卷曲组成,三级结构预测结果与其一致;Myoz1蛋白与ACTN2、LDB3、LMOD3等Z盘蛋白(PDZ-LIM结构域蛋白,ZASP)以及肌肉生长发育相关蛋白有相互作用的关系;Myoz1在1日龄大恒799肉鸡胸肌和腿肌的表达量显著高于其他组织,且在胸肌中的表达量高于腿肌(P<0.05)。研究表明Myoz1基因可能在肌肉生长发育过程中发挥调控作用。To study the amino acid sequence characteristics of calcineurin muscle nodular binding protein 1(Myoz1)and its mRNA expression level in different tissues and organs,the CDs sequence of Myoz1 was cloned by PCR.ExPASYProtparam,ProtScale,Target P,DNAStar Protean,SWISS-MODEL,SignalP 4.1 Server,NetPhos2.0Server and STRTNG software were re-spectively used to analyze the physicochemical properties,hydrophilicity/hydrophobicity,subcellular structure,protein secondary structure,tertiary structure,signal peptide,phosphorylation site and protein interaction of Myoz1 protein,and the expression level of Myoz1 gene was detected by real-time fluorescence quantitation PCR(qRT-PCR).The results showed that the total length of the Myoz1 gene CDs sequence in Daheng 799 broiler was 881 bp,which can encode 293 amino acids;The amino acid sequence of the Myoz1 gene in Daheng 799 broiler was located in the same branch as Phasianus colchicus and Lagopus muta,with the highest homology,both higher than 96%,and the closest genetic relationship;Myoz1 protein was water-soluble and mostly distributed in the nucleus.The molecular formula was C1442H_(2)244N400O430S10,the molecular weight was about 32384.70 ku,and the theoretical iso-electric point was 9.12;Myoz1 was a secreted protein without signal peptide,and had 35 potential phosphorylation sites and 1 po-tential N-glycosylation site;The secondary structure of Myoz1 protein was mainly composed ofα-helix,β-corner and random curl,which was consistent with the predicted results of the tertiary structure;Myoz1 interacted with Z-disk proteins such as ACTN2,LDB3,LMOD3(PDZ-LIM domain protein,ZASP)and other muscle growth and development related proteins;The ex-pression level of Myoz1 in breast muscle and leg muscle of 1-day-age Daheng 799 broiler was significantly higher than that in other tissues,and the expression level in chest muscle was higher than that in leg muscle(P<0.05).It was suggested that Myoz1 gene might play a regulatory role in muscle growth and development.

关 键 词：大恒肉鸡 Myoz1基因 克隆 生物信息学分析 组织表达 

分 类 号：S831.2[农业科学—畜牧学]