黄颡鱼硒蛋白基因的克隆与分析
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作者单位:

1.华中农业大学水产学院,武汉430070;2.华中农业大学深圳营养与健康研究院,武汉430070;3.中国农业科学院深圳农业基因组研究所/岭南现代农业科学与技术广东省实验室深圳分中心,深圳 518000

作者简介:

刘光辉,E-mail: ghliu@webmail.hzau.edu.cn

通讯作者:

罗智,E-mail: luozhi99@mail.hzau.edu.cn

中图分类号:

Q959.4

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)


Cloning and analysis of selenoprotein genes in yellow catfish,Pelteobagrus fulvidraco
Author:
Affiliation:

1.College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China;2.Shenzhen Institute of Nutrition and Health,Huazhong Agricultural University,Wuhan 430070,China;3.Shenzhen Branch,Guangdong Laboratory for Lingnan Modern Agriculture,Genome Analysis Laboratory of the Ministry of Agriculture/Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen 518000,China

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    摘要:

    为探讨黄颡鱼硒蛋白selenow2aselenop2selenot2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selenow2aselenop2selenot2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 selenow2aselenop2 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 selenocysteine 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 selenow2aselenop2 and selenot2 genes shared 82.24%,66.19%,and 79.45% similarity with those of zebrafish (Danio rerio),respectively,and shared 94.74%,68.50% and 90.95% similarity 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.

    表 3 硒蛋白基因cDNA序列信息Table 3 cDNA sequence information of the selenoprotein genes
    表 1 黄颡鱼硒蛋白基因cDNA克隆引物序列Table 1 Nucleotide sequences of the primers used for the cDNA cloning
    Fig.
    图1 黄颡鱼selenow2a基因(A)、selenop2基因(B)和selenot2基因(C)的核苷酸序列及其氨基酸序列和功能域Fig.1 Nucleotide sequence of the selenow2a(A),selenop2(B) and selenot2(C) gene of yellow catfish and its deduced amino acid sequence and predicted functional domain
    图2 Selenow2a(A)、Selenop2(B)和Selenot2(C)氨基酸序列多重比对Fig.2 Multiple amino acid sequence alignment of Selenow2a(A),Selenop2(B) and Selenot2(C)
    图3 黄颡鱼硒蛋白SECIS元件分析与比较Fig.3 Analysis and comparison of selenoprotein selenocysteine insertion sequence (SECIS)elements of selenoprotein genes from yellow catfish
    图4 Selenow2a(A)、Selenop2(B)和Selenot2(C)氨基酸序列系统进化树Fig.4 Phylogenetic tree based on Selenow2a (A),Selenop2 (B) and Selenot2 (C) sequences
    图5 硒蛋白基因selenow2a (A),selenop2 (B) 和selenot2 (C)在黄颡鱼不同组织中的分布和表达Fig.5 The distribution and expression of selenoprotein genes selenow2a (A),selenop2 (B) and selenot2 (C) in different tissues in yellow catfish
    表 2 硒蛋白基因荧光定量PCR引物Table 2 Primers for quantitative PCR of selenoprotein genes
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引用本文

刘光辉,余岸艮,何杨,郭雨诗,柯江,罗智.黄颡鱼硒蛋白基因的克隆与分析[J].华中农业大学学报,2024,43(1):185-193

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  • 收稿日期:2022-11-02
  • 在线发布日期: 2024-01-30
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