斑節(jié)對(duì)蝦含硒谷胱甘肽過(guò)氧化物酶基因全長(zhǎng)克隆及表達(dá)分析
發(fā)布時(shí)間：2018-11-08 20:05
【摘要】：為了研究含硒谷胱甘肽過(guò)氧化物酶(selenium-dependent glutathione peroxidase,SeGPx)在斑節(jié)對(duì)蝦應(yīng)激反應(yīng)和卵巢發(fā)育中的作用,實(shí)驗(yàn)以斑節(jié)對(duì)蝦肝胰腺轉(zhuǎn)錄組數(shù)據(jù)中篩選獲得的Se-GPx基因(Pm Se-GPx)片段為基礎(chǔ),利用RACE技術(shù)獲得Pm Se-GPx基因的c DNA全長(zhǎng),并對(duì)其進(jìn)行了生物信息學(xué)分析;利用熒光定量PCR技術(shù)研究了Pm Se-GPx在斑節(jié)對(duì)蝦不同組織、卵巢發(fā)育各期、肝胰腺組織在pH9、硫酸銅(Cu~(2+))應(yīng)激下的相對(duì)表達(dá)量和變化趨勢(shì)。結(jié)果顯示,Pm Se-GPx基因c DNA全長(zhǎng)為959 bp,5'非編碼區(qū)(UTR)長(zhǎng)10bp,開(kāi)放閱讀框(ORF)長(zhǎng)639 bp,編碼212個(gè)氨基酸,310 bp的3'UTR包含一個(gè)硒代半胱氨酸插入序列(SECIS),ORF中的密碼子209TGA211編碼硒代半胱氨酸(selenocysteine,U67)。實(shí)時(shí)定量PCR實(shí)驗(yàn)結(jié)果表明,Pm Se-GPx在斑節(jié)對(duì)蝦的肝胰腺、卵巢、精巢、心臟等組織中均有表達(dá),其中肝胰腺中表達(dá)量顯著高于其他組織,卵巢次之;卵巢發(fā)育階段表達(dá)結(jié)果則顯示Pm Se-GPx在卵巢發(fā)育各期均有表達(dá),在卵巢Ⅲ期表達(dá)量最高,其次是V期;在Cu~(2+)脅迫下,其表達(dá)量總體呈現(xiàn)先下降后上升然后又下降的趨勢(shì);在環(huán)境pH為9脅迫下,表達(dá)趨勢(shì)呈現(xiàn)先下降,后回升,然后又下降,再大幅上升的趨勢(shì)。研究表明,SeGPx在斑節(jié)對(duì)蝦卵巢發(fā)育過(guò)程中具有重要作用,且Se-GPx參與斑節(jié)對(duì)蝦對(duì)環(huán)境理化因子應(yīng)激的免疫調(diào)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，
本文編號(hào)：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，
本文編號(hào)：2319524
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