本文關(guān)鍵詞： 牙鲆 dazl cDNA克隆 基因表達(dá)　出處：《海洋漁業(yè)》2016年04期 　論文類型：期刊論文

【摘要】：dazl(deleted in azoospermia-like)基因是多種物種精子發(fā)生的重要調(diào)控因子,為探討該基因在牙鲆(Paralichthys olivaceus)性腺發(fā)育分化中的作用,本研究采用同源克隆和RACE技術(shù)克隆了牙鲆dazl基因的cDNA序列,利用生物信息學(xué)方法分析了其序列結(jié)構(gòu),并利用實(shí)時(shí)熒光定量PCR技術(shù)檢測了該基因在牙鲆成體組織和早期發(fā)育階段的表達(dá)情況。結(jié)果發(fā)現(xiàn):牙鲆dazl cDNA序列全長2 031 bp,包括105 bp的5'端非翻譯區(qū),1 275 bp的3'端非翻譯區(qū)和編碼217個(gè)氨基酸殘基的651 bp開放閱讀框;氨基酸同源序列比對顯示該序列存在一個(gè)高度保守的RRM(RNA recognition motif)結(jié)構(gòu)域,且該結(jié)構(gòu)域在魚類中具有100%的序列一致性;熒光定量PCR結(jié)果顯示dazl基因主要在牙鲆性腺中表達(dá),且在精巢中的表達(dá)高于卵巢;在牙鲆早期發(fā)育階段,dazl基因在未受精卵、受精卵及囊胚期均有較高的轉(zhuǎn)錄本,而在原腸胚期表達(dá)開始降低,至孵化后3 d一直保持較低的水平,這些豐富表達(dá)的母源性dazl mRNA可能參與了胚胎發(fā)育中原始生殖細(xì)胞的形成。本研究為進(jìn)一步闡明dazl基因在牙鲆生殖發(fā)育中的功能奠定了基礎(chǔ)。
[Abstract]:Dazl(deleted in azoospermia-like gene is an important regulator of spermatogenesis in many species. To investigate the role of this gene in the development and differentiation of gonad of Paralichthys olivaceus in Paralichthys olivaceus. In this study, the cDNA sequence of dazl gene of Paralichthys olivaceus was cloned by homologous cloning and RACE technique, and its sequence structure was analyzed by bioinformatics. The expression of the gene in the adult tissues and early development stage of Paralichthys olivaceus was detected by real-time fluorescence quantitative PCR. The results showed that the dazl cDNA sequence of Paralichthys olivaceus was 2031 BP in length. It includes a 5'untranslated region of 105 BP and a 3'terminal untranslated region of 1 275 BP and an open reading frame of 651 BP encoding 217 amino acid residues. The amino acid sequence alignment showed that the sequence had a highly conserved RRM(RNA recognition motif domain. The domain has a sequence consistency of 100% in fish. Fluorescence quantitative PCR showed that dazl gene was mainly expressed in the gonads of Paralichthys olivaceus and was higher in the testis than in the ovaries. In the early stage of development of Paralichthys olivaceus, there were high transcripts of dazl gene in unfertilized eggs, fertilized eggs and blastocysts, but the expression of dazl gene began to decrease at the stage of proembryo, and remained at a low level until 3 days after hatching. These abundant expression of maternal dazl. MRNA may be involved in the formation of primordial germ cells in embryonic development. This study lays a foundation for further elucidating the function of dazl gene in the reproductive development of Paralichthys olivaceus (Paralichthys olivaceus).
【作者單位】： 上海海洋大學(xué)水產(chǎn)與生命學(xué)院農(nóng)業(yè)部淡水水產(chǎn)種質(zhì)資源重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金青年項(xiàng)目(41306128);國家自然科學(xué)基金面上項(xiàng)目(30271017)
【分類號】：S917.4
【正文快照】： DAZ基因家族包括daz、dazl(daz-like)和bolue基因,這3者的編碼蛋白均為RNA結(jié)合蛋白,在生殖細(xì)胞中特異表達(dá),參與調(diào)節(jié)生殖細(xì)胞的發(fā)育和分化[1-2],是精子發(fā)生過程中減數(shù)分裂的主要調(diào)控因子[3]。bolue和dazl是常染色體基因,一般認(rèn)為bolue是DAZ基因家族的祖先基因(ancestral gene),

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