斑节对虾含硒谷胱甘肽过氧化物酶基因全长克隆及表达分析
发布时间：2018-11-08 20:05
【摘要】：为了研究含硒谷胱甘肽过氧化物酶(selenium-dependent glutathione peroxidase,SeGPx)在斑节对虾应激反应和卵巢发育中的作用,实验以斑节对虾肝胰腺转录组数据中筛选获得的Se-GPx基因(Pm Se-GPx)片段为基础,利用RACE技术获得Pm Se-GPx基因的c DNA全长,并对其进行了生物信息学分析;利用荧光定量PCR技术研究了Pm Se-GPx在斑节对虾不同组织、卵巢发育各期、肝胰腺组织在pH9、硫酸铜(Cu~(2+))应激下的相对表达量和变化趋势。结果显示,Pm Se-GPx基因c DNA全长为959 bp,5'非编码区(UTR)长10bp,开放阅读框(ORF)长639 bp,编码212个氨基酸,310 bp的3'UTR包含一个硒代半胱氨酸插入序列(SECIS),ORF中的密码子209TGA211编码硒代半胱氨酸(selenocysteine,U67)。实时定量PCR实验结果表明,Pm Se-GPx在斑节对虾的肝胰腺、卵巢、精巢、心脏等组织中均有表达,其中肝胰腺中表达量显著高于其他组织,卵巢次之;卵巢发育阶段表达结果则显示Pm Se-GPx在卵巢发育各期均有表达,在卵巢Ⅲ期表达量最高,其次是V期;在Cu~(2+)胁迫下,其表达量总体呈现先下降后上升然后又下降的趋势;在环境pH为9胁迫下,表达趋势呈现先下降,后回升,然后又下降,再大幅上升的趋势。研究表明,SeGPx在斑节对虾卵巢发育过程中具有重要作用,且Se-GPx参与斑节对虾对环境理化因子应激的免疫调控。
[Abstract]:To study the role of selenium-containing glutathione peroxidase (selenium-dependent glutathione peroxidase,SeGPx) in stress response and ovarian development of Penaeus Monodon. Based on the Se-GPx gene (Pm Se-GPx) fragment selected from the transcriptome data of the hepatopancreas of Penaeus Monodon (Penaeus Monodon), the full length of c DNA of Pm Se-GPx gene was obtained by RACE technique and analyzed by bioinformatics. The relative expression of Pm Se-GPx in different tissues of Penaeus Monodon and in different stages of ovary development and in hepatopancreas under the stress of pH9, (Cu~ (2) was studied by fluorescence quantitative PCR technique. The results showed that the length of c DNA of Pm Se-GPx gene was 959 bp,5', the length of (UTR) was 10 BP, the length of open reading frame (ORF) 639 bp, encoded 212 amino acids, and the 3'UTR of 310 bp contained a selenocysteine insertion sequence (SECIS),. The codon 209TGA211 in ORF encodes selenocysteine (selenocysteine,U67). The results of real-time quantitative PCR showed that Pm Se-GPx was expressed in hepatopancreas, ovary, testis, heart and other tissues of Penaeus Monodon. The expression of Pm Se-GPx in hepatopancreas was significantly higher than that in other tissues, followed by ovary. The expression of Pm Se-GPx was found in all stages of ovarian development, the highest in stage 鈪，
本文编号：2319524
[Abstract]:To study the role of selenium-containing glutathione peroxidase (selenium-dependent glutathione peroxidase,SeGPx) in stress response and ovarian development of Penaeus Monodon. Based on the Se-GPx gene (Pm Se-GPx) fragment selected from the transcriptome data of the hepatopancreas of Penaeus Monodon (Penaeus Monodon), the full length of c DNA of Pm Se-GPx gene was obtained by RACE technique and analyzed by bioinformatics. The relative expression of Pm Se-GPx in different tissues of Penaeus Monodon and in different stages of ovary development and in hepatopancreas under the stress of pH9, (Cu~ (2) was studied by fluorescence quantitative PCR technique. The results showed that the length of c DNA of Pm Se-GPx gene was 959 bp,5', the length of (UTR) was 10 BP, the length of open reading frame (ORF) 639 bp, encoded 212 amino acids, and the 3'UTR of 310 bp contained a selenocysteine insertion sequence (SECIS),. The codon 209TGA211 in ORF encodes selenocysteine (selenocysteine,U67). The results of real-time quantitative PCR showed that Pm Se-GPx was expressed in hepatopancreas, ovary, testis, heart and other tissues of Penaeus Monodon. The expression of Pm Se-GPx in hepatopancreas was significantly higher than that in other tissues, followed by ovary. The expression of Pm Se-GPx was found in all stages of ovarian development, the highest in stage 鈪，
本文编号：2319524
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