【摘要】：以仿刺參(Apostichopus japonicus)表觀遺傳調控相關基因—DNA甲基轉移酶(DNA methyltransferase,DNMT1)、組蛋白去乙�；�(histone deacetylase,HDAC3)和組蛋白甲基轉移酶(histone methyltransferase,MLL5)為目的基因,采用實時熒光定量PCR(q RT-PCR)技術和相對定量2~(-ΔΔC_T)法,以刺參管家基因β-actin為內參進行校正,比較了高溫脅迫下3個基因在呼吸樹、消化道、體液、肌肉各組織中的表達情況。以15℃條件下的刺參為對照組,其目的基因表達量為基準1,在體液中,DNMT1基因在18℃時表達量上升不顯著(P0.05),18~21℃時表達量上升極顯著(P0.01),21~24℃時上升不明顯(P0.05),30℃得到最大表達量為3.26,總體為上調表達;HDAC3基因和MLL5基因在24℃時的表達量顯著(P0.05),之后隨著溫度升高其表達量稍有升高,分別在30℃得到最大表達量為2.90和3.19�？傮w來看,各基因隨著溫度的升高其表達量都增高,上調表達趨勢明顯,在30℃達到最高表達量。對比3個基因在不同組織中的表達差異,體液中表達變化較明顯,在24℃以后均大于2.5;肌肉中最低,在整個高溫脅迫過程中基因表達變化量小于2。在30℃的溫度脅迫條件下,檢測在0 h、3 h、6 h、9 h、12 h、15 h 6個時間點3個基因的表達量,以0 h各基因的表達量為基準1,結果顯示,3個基因的表達量都表現(xiàn)為上調表達,除消化道的HDAC3在6 h時提前進入平穩(wěn)期,其他各組織各基因的表達量均在0~9 h內隨著脅迫時間的延長呈現(xiàn)快速上升狀態(tài)(P0.05);脅迫9 h以后,各目的基因表達量變化不明顯或稍有降低(P0.05)。不同組織中目的基因的表達趨勢一致,體液中3種表觀相關基因的表達量最高值高于其他3種組織。本研究為從表觀遺傳修飾角度解析刺參的高溫逆境應答反應機制奠定了基礎。
[Abstract]:The target genes were DNA methyltransferase (DNA methyltransferase,DNMT1), histone deacetylase (histone deacetylase,HDAC3) and histone methyltransferase (histone methyltransferase,MLL5), which were related to (Apostichopus japonicus) epigenetic regulation. Real time fluorescence quantitative PCR (q RT-PCR) technique and relative quantitative 2 ~ (- 螖 C\ +\ {T\} T) method were used to calibrate the housekeeping gene 尾-actin of sea cucumber. Three genes were compared in respiratory tree, digestive tract and body fluid under high temperature stress. Expression in muscle tissues. The target gene expression of Acanthopsis japonicus under the condition of 15 鈩，

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