【摘要】：本研究應(yīng)用RACE技術(shù)克隆了青蝦(Macrobrachium nipponensis)Ran(Ras related nuclear protein,Ras相關(guān)核蛋白)基因全長cDNA序列,該基因cDNA全長1191 bp,包括218 bp的5'UTR,648 bp的開放閱讀框(ORF),405 bp的3'UTR,編碼215個(gè)氨基酸。青蝦Ran基因?qū)儆赑-loop-NTPase超級家族,擁有PTZ00132跨結(jié)構(gòu)域,多肽分子量約為24.57 kDa,理論等電點(diǎn)7.13。系統(tǒng)進(jìn)化樹分析表明,在動(dòng)物界進(jìn)化中非常保守的青蝦Ran多肽與羅氏沼蝦(Macrobrachium rosenbergii)聚為一支,具有最近的親緣關(guān)系。使用熒光定量PCR技術(shù)檢測Ran基因在成體青蝦不同組織和卵巢不同發(fā)育期的表達(dá)差異,結(jié)果顯示,Ran基因在青蝦不同組織中均有表達(dá),其表達(dá)量在卵巢中最高,是精巢表達(dá)量的7~8倍;隨著卵巢的發(fā)育,Ran基因的表達(dá)水平呈現(xiàn)上升趨勢,在卵巢消退期又恢復(fù)到較低水平。RNA干擾后,實(shí)驗(yàn)組Ran基因表達(dá)量顯著低于對照組(P0.05),同時(shí)卵巢發(fā)育關(guān)鍵基因Vg(vitellogenin)在卵巢中的表達(dá)量也顯著低于對照組(P0.05),推測Ran基因參與雌性卵巢發(fā)育過程并對Vg基因的表達(dá)起到調(diào)控作用。
[Abstract]:In this study, the full-length cDNA sequence of the shrimp (Macrobrachium nipponensis) Ran (Ras related nuclear protein,Ras-associated nucleoprotein (Macrobrachium nipponensis) Ran (Ras related nuclear protein,Ras-associated nucleoprotein) gene was cloned by RACE technique. The full-length cDNA gene consists of 5 UTRs of 218 bp and 3 UTRs of (ORF), 405bp of the open reading frame (ORF) of 648 bp. It encodes 215 amino acids. The Ran gene of shrimps belongs to the P-loop-NTPase superfamily and has a PTZ00132 domain. The molecular weight of the polypeptide is about 24.57 kDa, theoretical isoelectric point 7.13. Phylogenetic tree analysis showed that the very conservative Ran polypeptide of Macrobrachium rosenbergii and Macrobrachium rosenbergii (Macrobrachium rosenbergii) were closely related to each other in the evolution of animal kingdom. The expression of Ran gene was detected by fluorescence quantitative PCR in different tissues of adult shrimps and in different developmental stages of ovary. The results showed that Ran gene was expressed in different tissues of shrimps, and the expression level of Ran gene was the highest in ovary. It was 8 times higher than that of testis. With the development of ovary, the expression level of Ran gene showed an increasing trend, and returned to a lower level in the ovariectomized period. After RNA interference, the expression of Ran gene in the experimental group was significantly lower than that in the control group (P0.05). At the same time, the expression of Vg (vitellogenin), the key gene of ovarian development, was significantly lower than that of the control group (P0.05). It was speculated that Ran gene was involved in the development of female ovary and regulated the expression of Vg gene.
【作者單位】： 南京農(nóng)業(yè)大學(xué)無錫漁業(yè)學(xué)院;中國水產(chǎn)科學(xué)研究院淡水漁業(yè)研究中心農(nóng)業(yè)部淡水漁業(yè)和種質(zhì)資源利用重點(diǎn)實(shí)驗(yàn)室;
【基金】：江蘇省重點(diǎn)研發(fā)計(jì)劃(現(xiàn)代農(nóng)業(yè))重點(diǎn)項(xiàng)目(BE2016308) 中國水產(chǎn)科學(xué)研究院基本科研業(yè)務(wù)費(fèi)專項(xiàng)項(xiàng)目(2016HY-ZD0402) 國家自然科學(xué)基金面上項(xiàng)目(31572617) 江蘇省農(nóng)業(yè)科技自主創(chuàng)新基金項(xiàng)目[CX(15)10124] 江蘇省水產(chǎn)三新工程(D2015-16) 無錫科學(xué)科技發(fā)展基金項(xiàng)目(CLE02N1514)
【分類號】：S917.4
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本文編號：2444911