【摘要】：為研究冷休克蛋白Y-box基因在青蝦應答環(huán)境脅迫過程中所起的調(diào)控作用,實驗應用RACE PCR技術首次克隆了青蝦的冷休克蛋白Y-box基因全長c DNA序列,并利用在線軟件對其序列特征進行生物信息學分析;采用實時熒光定量PCR技術對其在青蝦不同組織及環(huán)境脅迫過程中的表達變化特征進行分析。青蝦冷休克蛋白Y-box基因c DNA全長1501 bp,包括84 bp的5′末端非翻譯區(qū)(UTR),876 bp的開放閱讀框(ORF),541 bp的3′UTR,開放閱讀框編碼291個氨基酸。氨基酸相似度比對顯示,青蝦冷休克蛋白Y-box基因富含高度保守的冷休克結(jié)構域。系統(tǒng)進化樹分析顯示,青蝦冷休克蛋白Y-box基因與水蚤等節(jié)肢動物冷休克Y-box聚類一支,具有最近的親緣關系。熒光定量PCR檢測顯示,冷休克蛋白Y-box基因在青蝦不同組織中均有表達,其表達量在肝胰腺組織中最高,使用熒光定量PCR檢測青蝦冷休克蛋白Y-box基因在低溫脅迫和恢復條件下在肝胰腺中的m RNA時空表達情況,結(jié)果顯示,與對照組相比冷休克蛋白Y-box在肝胰腺中的表達量分別在低溫和低氧脅迫3,6和12 h出現(xiàn)了顯著上調(diào),而在恢復刺激后其表達量與對照組差異不顯著。此外,本實驗對Y-box進行了原核表達,為進一步研究Y-box基因的功能奠定了基礎。
[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.
【作者單位】： 中國水產(chǎn)科學研究院淡水漁業(yè)研究中心農(nóng)業(yè)部淡水漁業(yè)與種質(zhì)資源利用重點實驗室;南京農(nóng)業(yè)大學無錫漁業(yè)學院;
【基金】：國家自然科學基金(31402280,31672633) 國家“十二五”科技支撐計劃(2012BAD25B07)~~
【分類號】：S917.4
，

本文編號：2452483