【摘要】：以仿刺參(Apostichopus japonicus)表觀遺傳調(diào)控相關(guān)基因—DNA甲基轉(zhuǎn)移酶(DNA methyltransferase,DNMT1)、組蛋白去乙�；�(histone deacetylase,HDAC3)和組蛋白甲基轉(zhuǎn)移酶(histone methyltransferase,MLL5)為目的基因,采用實(shí)時(shí)熒光定量PCR(q RT-PCR)技術(shù)和相對(duì)定量2~(-ΔΔC_T)法,以刺參管家基因β-actin為內(nèi)參進(jìn)行校正,比較了高溫脅迫下3個(gè)基因在呼吸樹、消化道、體液、肌肉各組織中的表達(dá)情況。以15℃條件下的刺參為對(duì)照組,其目的基因表達(dá)量為基準(zhǔn)1,在體液中,DNMT1基因在18℃時(shí)表達(dá)量上升不顯著(P0.05),18~21℃時(shí)表達(dá)量上升極顯著(P0.01),21~24℃時(shí)上升不明顯(P0.05),30℃得到最大表達(dá)量為3.26,總體為上調(diào)表達(dá);HDAC3基因和MLL5基因在24℃時(shí)的表達(dá)量顯著(P0.05),之后隨著溫度升高其表達(dá)量稍有升高,分別在30℃得到最大表達(dá)量為2.90和3.19。總體來(lái)看,各基因隨著溫度的升高其表達(dá)量都增高,上調(diào)表達(dá)趨勢(shì)明顯,在30℃達(dá)到最高表達(dá)量。對(duì)比3個(gè)基因在不同組織中的表達(dá)差異,體液中表達(dá)變化較明顯,在24℃以后均大于2.5;肌肉中最低,在整個(gè)高溫脅迫過(guò)程中基因表達(dá)變化量小于2。在30℃的溫度脅迫條件下,檢測(cè)在0 h、3 h、6 h、9 h、12 h、15 h 6個(gè)時(shí)間點(diǎn)3個(gè)基因的表達(dá)量,以0 h各基因的表達(dá)量為基準(zhǔn)1,結(jié)果顯示,3個(gè)基因的表達(dá)量都表現(xiàn)為上調(diào)表達(dá),除消化道的HDAC3在6 h時(shí)提前進(jìn)入平穩(wěn)期,其他各組織各基因的表達(dá)量均在0~9 h內(nèi)隨著脅迫時(shí)間的延長(zhǎng)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(dá)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(dá)趨勢(shì)一致,體液中3種表觀相關(guān)基因的表達(dá)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(yīng)答反應(yīng)機(jī)制奠定了基礎(chǔ)。
[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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