青虾冷休克蛋白Y-box编码基因的cDNA全长克隆与表达分析
发布时间：2019-04-02 10:26
【摘要】：为研究冷休克蛋白Y-box基因在青虾应答环境胁迫过程中所起的调控作用,实验应用RACE PCR技术首次克隆了青虾的冷休克蛋白Y-box基因全长c DNA序列,并利用在线软件对其序列特征进行生物信息学分析;采用实时荧光定量PCR技术对其在青虾不同组织及环境胁迫过程中的表达变化特征进行分析。青虾冷休克蛋白Y-box基因c DNA全长1501 bp,包括84 bp的5′末端非翻译区(UTR),876 bp的开放阅读框(ORF),541 bp的3′UTR,开放阅读框编码291个氨基酸。氨基酸相似度比对显示,青虾冷休克蛋白Y-box基因富含高度保守的冷休克结构域。系统进化树分析显示,青虾冷休克蛋白Y-box基因与水蚤等节肢动物冷休克Y-box聚类一支,具有最近的亲缘关系。荧光定量PCR检测显示,冷休克蛋白Y-box基因在青虾不同组织中均有表达,其表达量在肝胰腺组织中最高,使用荧光定量PCR检测青虾冷休克蛋白Y-box基因在低温胁迫和恢复条件下在肝胰腺中的m RNA时空表达情况,结果显示,与对照组相比冷休克蛋白Y-box在肝胰腺中的表达量分别在低温和低氧胁迫3,6和12 h出现了显著上调,而在恢复刺激后其表达量与对照组差异不显著。此外,本实验对Y-box进行了原核表达,为进一步研究Y-box基因的功能奠定了基础。
[Abstract]:In order to study the regulation of cold shock protein Y-box gene in response to environmental stress, the full length c DNA sequence of cold shock protein Y-box gene was cloned by RACE PCR technique for the first time. On-line software was used for bioinformatics analysis of its sequence characteristics. Real-time fluorescence quantitative PCR (real-time quantitative PCR) was used to analyze the characteristics of its expression in different tissues and environmental stresses of shrimps. The full length of c DNA of cold shock protein Y-box gene of shrimps is 1501 bp, including the open reading frame (ORF) of (ORF), 541 bp of the untranslated region (UTR), 876 bp of 84 bp. The open reading frame encodes 291 amino acids. Amino acid similarity ratio showed that the cold shock protein Y-box gene of shrimps was rich in highly conserved cold shock domains. Phylogenetic tree analysis showed that the cold shock protein Y-box gene of shrimps was closely related to the cold shock Y-box of arthropods such as water fleas. Fluorescence quantitative PCR analysis showed that cold shock protein Y-box gene was expressed in different tissues of shrimps, and its expression level was the highest in hepatopancreas. The spatial and temporal expression of cold shock protein Y-box gene in hepatopancreas of shrimps was detected by fluorescence quantitative PCR under low temperature stress and recovery. The results showed that the expression of cold shock protein Y-box gene in hepatopancreas was observed. Compared with the control group, the expression of cold shock protein Y-box in hepatopancreas was up-regulated at 3 h, 6 h and 12 h respectively under hypothermia and hypoxia stress, but there was no significant difference between the control group and the control group after the recovery of stimulation. In addition, the prokaryotic expression of Y-box laid a foundation for further study on the function of Y-box gene.
【作者单位】： 中国水产科学研究院淡水渔业研究中心农业部淡水渔业与种质资源利用重点实验室;南京农业大学无锡渔业学院;
【基金】：国家自然科学基金(31402280,31672633) 国家“十二五”科技支撑计划(2012BAD25B07)~~
【分类号】：S917.4
，
本文编号：2452483
[Abstract]:In order to study the regulation of cold shock protein Y-box gene in response to environmental stress, the full length c DNA sequence of cold shock protein Y-box gene was cloned by RACE PCR technique for the first time. On-line software was used for bioinformatics analysis of its sequence characteristics. Real-time fluorescence quantitative PCR (real-time quantitative PCR) was used to analyze the characteristics of its expression in different tissues and environmental stresses of shrimps. The full length of c DNA of cold shock protein Y-box gene of shrimps is 1501 bp, including the open reading frame (ORF) of (ORF), 541 bp of the untranslated region (UTR), 876 bp of 84 bp. The open reading frame encodes 291 amino acids. Amino acid similarity ratio showed that the cold shock protein Y-box gene of shrimps was rich in highly conserved cold shock domains. Phylogenetic tree analysis showed that the cold shock protein Y-box gene of shrimps was closely related to the cold shock Y-box of arthropods such as water fleas. Fluorescence quantitative PCR analysis showed that cold shock protein Y-box gene was expressed in different tissues of shrimps, and its expression level was the highest in hepatopancreas. The spatial and temporal expression of cold shock protein Y-box gene in hepatopancreas of shrimps was detected by fluorescence quantitative PCR under low temperature stress and recovery. The results showed that the expression of cold shock protein Y-box gene in hepatopancreas was observed. Compared with the control group, the expression of cold shock protein Y-box in hepatopancreas was up-regulated at 3 h, 6 h and 12 h respectively under hypothermia and hypoxia stress, but there was no significant difference between the control group and the control group after the recovery of stimulation. In addition, the prokaryotic expression of Y-box laid a foundation for further study on the function of Y-box gene.
【作者单位】： 中国水产科学研究院淡水渔业研究中心农业部淡水渔业与种质资源利用重点实验室;南京农业大学无锡渔业学院;
【基金】：国家自然科学基金(31402280,31672633) 国家“十二五”科技支撑计划(2012BAD25B07)~~
【分类号】：S917.4
，
本文编号：2452483
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