摘要
为探讨黄颡鱼硒蛋白selenow2a、selenop2和selenot2基因之间的关系,采用3′/5′ RACE PCR克隆得到3个基因的cDNA全长,分别为891、1 998和1 432 bp,其中ORF长度分别为288、828和600 bp,编码95、275和219个氨基酸.在线工具SECISerach3对3个基因的cDNA序列分析结果显示,它们都含有可以编码硒代半胱氨酸的终止密码子,以及在3′非编码区存在SECIS元件.通过氨基酸序列比对和系统发育树分析,发现sele-now2a、selenop2和selenot2基因预测得到的氨基酸序列与斑马鱼(Danio rerio)氨基酸相似性分别为82.24%、66.19%和79.45%,而与斑点叉尾鮰(Ictalurus punetaus)的氨基酸相似性分别为94.74%、68.50%和90.95%,在发育树上则显示为树杈相接近.采用实时荧光定量PCR检测3个硒蛋白基因的mRNA在黄颡鱼心脏、肝脏、肌肉、脑、肠、脾脏、精巢和卵巢组织中的表达,结果显示其mRNA表达水平呈现组织特异性.表明3个基因拥有硒蛋白家族的特征,但在组织表达上具有特异性.
Abstract
In this study,to investigate the relationship among the three selenoprotein genes sele-now2a,selenop2 and selenot2 in yellow catfish,Pelteobagrus fulvidraco,3′/5′ RACE PCR was used to clone the full length cDNAs of three genes,which were 891,1 998 and 1 432 bp with the ORFs of 288,828 and 600 bp,encoding 95,275 and 219 amino acids,respectively.The cDNA sequence analysis of the three genes by online tool SECISerach3 showed that they all contained the stop codons encoding selenocys-teine and the selenocysteine insertion sequence(SECIS)element in the 3′ untranslated region(3′ UTR).Through amino acid sequence comparison and phylogenetic tree analysis,it was found that the predicted amino acid sequences of selenow2a,selenop2 and selenot2 genes shared 82.24%,66.19%,and 79.45%simi-larity with those of zebrafish(Danio rerio),respectively,and shared 94.74%,68.50%and 90.95%similari-ty with those of channel catfish(Ictalurus punetaus),respectively,which was shown to be close to the fork of the tree.Quantitative real-time PCR was used to examine the mRNA expression of three selenoprotein genes in the heart,liver,muscle,brain,intestine,spleen,testis and ovaries of yellow catfish,and the results suggested that the mRNA expression levels were tissue-specific.These results suggested that these three genes share the characteristics of selenoprotein family,but are tissue-specific in expression.
基金项目
华中农业大学深圳营养与健康研究院项目(SZYJY2023016)
国家重点研发计划"蓝色粮仓"科技创新专项(2018YFD0900400)
NETL
NSTL
维普
万方数据