牛源CCL28基因克隆、生物学特性及mRNA表达分析  

Cloning,biological characteristics and mRNA expression analysis of CCL28 gene in bovine

作　　者：姚竞杰 徐萍[1,2] 陈伶慧 申祥 白跃宇 王三虎 张晓建[1] YAO Jingjie;XU Ping;CHEN Linghui;SHEN Xiang;BAI Yueyu;WANG Sanhu;ZHANG Xiaojian(College of Animal Science and Technology,Henan Institute of Science and Technology,Xinxiang 453003,China;Department of Veterinary Medicine,Sumy National Agrarian University,Sumy 40021,Ukraine;Animal Health Supervision of Henan Province,Zhengzhou 410100,China)

机构地区：[1]河南科技学院动物科技学院,河南新乡453003 [2]苏梅国立农业大学兽医学院,乌克兰苏梅40021 [3]河南省动物卫生监督所,郑州410100

出　　处：《黑龙江畜牧兽医》2021年第9期12-16,161,162,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：河南省基础与前沿技术研究计划项目(152300410095);NSFC-河南人才培养联合基金项目(U1504306);2014年度高层次人才科研启动项目(206010615007)。

摘　　要：为了探究牛趋化因子CCL28基因编码蛋白的生物学特性及其mRNA在奶牛不同器官中的表达水平,试验采用PCR方法扩增并克隆奶牛CCL28基因CDS全长序列;通过生物信息学软件对其编码蛋白进行分析并构建系统进化树;应用实时荧光定量PCR方法检测CCL28基因mRNA在奶牛不同器官中的表达情况。结果表明:CCL28基因编码蛋白是一种结构较不稳定、带正电的亲水性蛋白,由129个氨基酸组成;牛CCL28基因核苷酸序列与绵羊的同源性较高,达到94.3%,与绵羊的遗传距离较近;CCL28蛋白的二级结构主要由无规则卷曲和α-螺旋构成,主要存在于细胞外,可能与CCR10、CCR3、ACKR2等蛋白存在互作关系;平均信号肽分值为0.901,没有跨膜结构域;CCL28基因mRNA在奶牛多个器官中均有表达,其中乳腺中的表达量极显著高于其他器官(P<0.01),肝脏和肾脏次之。说明CCL28基因可能在乳腺组织免疫过程中发挥调控作用。In order to explore the biological characteristics of the protein encoded by bovine chemokine CCL28 gene and its mRNA expression level in different tissues of dairy cows,the full-length CDS sequence of CCL28 gene of dairy cow was amplified and cloned by PCR.The protein was analyzed by bioinformatics software and phylogenetic tree was constructed.Real-time fluorescent quantitative PCR technology was used to detect the expression of CCL28 gene in different organs of dairy cows.The results showed that the protein encoded by the CCL28 gene was a hydrophilic protein with positive charge and unstable structure,which consisted of 129 amino acids.The nucleotide sequence of bovine CCl28 gene was 94.3%homologous to goats,which was close to that of sheep.The secondary structure of CCl28 protein was mainly composed of random coiling andα-helix,which mainly existed extracellular and might interact with CCR10,CCR3,ACKR2 and other proteins.The average signal peptide score was 0.901,and there was no transmembrane domain.CCL28 gene was expressed in many organs of dairy cows,and the expression level of CCL28 mRNA was higher in mammary gland than in other organs(P<0.01),followed by liver and kidney.It indicated that CCL28 gene might play a regulatory role in immunoregulation of mammary gland.

关 键 词：牛 CCL28基因 克隆 生物信息学分析 mRNA表达分析 

分 类 号：S823[农业科学—畜牧学] Q785[农业科学—畜牧兽医]