【摘要】：低氧是水產(chǎn)養(yǎng)殖動物面臨的主要脅迫之一。鳙(Hypophthalmichthys nobilis)是我國最重要的淡水養(yǎng)殖魚類之一,其對低氧脅迫敏感,但相關(guān)的遺傳基礎(chǔ)研究報道很少。本研究利用cDNA-擴(kuò)增片段長度多態(tài)性(cDNA-amplified fragment length polymorphism,cDNA-AFLP)技術(shù)對低氧敏感和低氧耐受兩組魚進(jìn)行比較分析,鑒定鳙與低氧反應(yīng)相關(guān)的信號通路中的差異表達(dá)基因。從Eco RⅠ和MseⅠAFLP體系的256個引物組合共獲得10 571條轉(zhuǎn)錄衍生片段(transcript-derived fragments,TDFs)。其中,低氧脅迫后差異表達(dá)的TDFs數(shù)目為221個,包括上調(diào)表達(dá)137個(62%),下調(diào)表達(dá)84個(38%)。對70個差異TDFs成功地進(jìn)行了切膠回收、PCR產(chǎn)物克隆和測序,序列比對分析后發(fā)現(xiàn):29個TDFs在數(shù)據(jù)庫中未檢索到同源序列;9個TDFs比對到未知功能序列;32個TDFs比對到已知生物學(xué)功能的序列,其功能包括轉(zhuǎn)錄調(diào)控、應(yīng)激和信號轉(zhuǎn)導(dǎo)、能量代謝、蛋白質(zhì)合成、免疫防御以及細(xì)胞生長與增殖。隨機(jī)選取4個已知功能的TDFs(ANXA6、TRIM25、Tnp和RBAT)進(jìn)行qRT-PCR驗證,結(jié)果顯示:ANXA6、TRIM25和Tnp在低氧脅迫后的表達(dá)模式與AFLP分析結(jié)果一致,RBAT與AFLP結(jié)果不一致。本研究結(jié)果有助于揭示鳙應(yīng)對低氧逆境的生理和遺傳調(diào)控機(jī)制,同時對鳙抗逆品種的分子標(biāo)記輔助選育也有一定的參考價值。
[Abstract]:Hypoxia is one of the main stresses faced by aquaculture animals. Bighead carp (Hypophthalmichthys nobilis) is one of the most important freshwater cultured fish in China. In this study, cDNA- amplified fragment length polymorphism (cDNA-amplified fragment length polymorphism,cDNA-AFLP) was used to compare the two groups of hypoxia sensitive and hypoxic tolerance, and to identify the differentially expressed genes in the signal pathway associated with hypoxia response of bighead carp (bighead carp). A total of 10 571 transcriptional derivative fragments (transcript-derived fragments,TDFs) were obtained from 256 primer combinations of Eco R 鈪，

本文編號：2206360