本文關(guān)鍵詞： 毛果楊 組蛋白去乙�；� 基因克隆 載體構(gòu)建 遺傳轉(zhuǎn)化 抗寒性　出處：《東北林業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：組蛋白去乙酰化酶(histone deacetylases, HDACs)催化組蛋白去乙�；�,參與組蛋白乙酰化狀態(tài)的調(diào)節(jié),對染色體結(jié)構(gòu)修飾和基因表達調(diào)控發(fā)揮著重要的作用。為了研究毛果楊HDA902基因的功能,采用PCR的方法克隆了毛果楊組蛋白去乙酰化酶基因HDA902的編碼序列,利用生物信息學(xué)方法分析了該基因所編碼的蛋白質(zhì),并構(gòu)建了GFP-HDA902融合基因表達載體,通過其在洋蔥表皮細胞的表達來確定HDA902蛋白的亞細胞定位。研究結(jié)果表明：(1)毛果楊HDA902基因的開放閱讀框為1500 bp,編碼一個由499個氨基酸殘基組成的蛋白質(zhì)。(2)毛果楊HDA902蛋白與其他植物同源蛋白具有一個保守的結(jié)構(gòu)域HDAC3。毛果楊HDA902蛋白在進化上與桃(Prunus persica)HDAC(XP 007200978.1)和蘋果(Malus domestica)HDA19-like(XP 008348955.1)的親緣關(guān)系較近。(3)毛果楊HDA902基因的啟動子序列包含5UTR Py-rich stretch、ARE、ACE和HSE等多個與逆境和光響應(yīng)相關(guān)的順式作用元件。(4)毛果楊HDA902蛋白主要定位于細胞質(zhì)中。(5)HDA902基因在毛果楊葉、莖和根三個組織中均有表達,但在莖和葉中表達量較高,根中的表達量相對較低。HDA902基因的表達受非生物脅迫的調(diào)節(jié),鹽脅迫降低了HDA902基因的表達,而低溫脅迫誘導(dǎo)了HDA902基因的表達。(6)構(gòu)建植物過量表達載體pROK Ⅱ-HDA902,通過農(nóng)桿菌介導(dǎo)法遺傳轉(zhuǎn)化到煙草中,對其抗逆性功能進行分析發(fā)現(xiàn),毛果楊HDA902基因在煙草中的表達提高了轉(zhuǎn)基因植株的抗寒性。
[Abstract]:Histone deacetylases (HDACs) catalyze histone deacetylation, participate in the regulation of histone acetylation state, and play an important role in chromosome structure modification and gene expression regulation. The coding sequence of histone deacetylase gene (HDA902) of Populus tomentosa was cloned by PCR. The protein encoded by this gene was analyzed by bioinformatics, and the expression vector of GFP-HDA902 fusion gene was constructed. The subcellular localization of HDA902 protein was determined by its expression in onion epidermis cells. The results showed that the open reading frame of HDA902 gene of Populus tomentosa was 1500 BP, encoding a 499-amino acid residue protein. HDA902 protein has a conserved domain with other plant homologous proteins, HDAC3.The phylogenetic relationship of HDA902 protein with Prunus persica)HDAC(XP 0077978.1) and Malus domestica)HDA19-like(XP 008348955.1) is closer.; the promoter sequence of HDA902 gene of Populus tomentosa contains 5UTR Py-rich stretchard ACE and ACE. HSE and other cis-acting elements related to stress and light response. 4) the HDA902 protein of Populus tomentosa was mainly located in the cytoplasm of P. hirsutum HDA902 gene in poplar leaves. The expression of HDA902 gene in stem and root was lower than that in stem and leaf, and the expression of HDA902 gene was regulated by abiotic stress. Salt stress decreased the expression of HDA902 gene. The expression of HDA902 gene was induced by low temperature stress. PROK 鈪，

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