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蒙古沙冬青J蛋白基因AmJ20的克隆與表達(dá)分析

發(fā)布時(shí)間:2018-09-18 20:17
【摘要】:DNA J蛋白是植物中廣泛存在的一類分子伴侶,對(duì)于維持正常和逆境條件下蛋白質(zhì)的穩(wěn)態(tài)具有重要作用。我們?cè)谇捌谕ㄟ^(guò)對(duì)強(qiáng)抗逆植物蒙古沙冬青進(jìn)行轉(zhuǎn)錄組分析,發(fā)現(xiàn)一個(gè)寒旱誘導(dǎo)表達(dá)的J蛋白即Am J20編碼序列,但其3'端編碼區(qū)不完整。本研究通過(guò)3'RACE獲得其c DNA全長(zhǎng)序列(800 bp),推測(cè)其編碼蛋白由179個(gè)氨基酸殘基組成,含一個(gè)典型的J結(jié)構(gòu)域,屬于Ⅲ型J蛋白,此外還具有葉綠體和細(xì)胞核雙定位信號(hào);利用半定量RT-PCR方法檢測(cè)該基因在野外生長(zhǎng)植株中的時(shí)空表達(dá)模式,發(fā)現(xiàn)其在春、夏、秋季只有微量表達(dá),甚至檢測(cè)不到轉(zhuǎn)錄本,而在嚴(yán)冬時(shí)節(jié)高表達(dá),此外在葉片中的表達(dá)量明顯高于根、嫩枝、花蕾和幼嫩果莢;利用RT-PCR方法克隆到該基因編碼區(qū)c DNA并將其插入到植物超表達(dá)載體上,為通過(guò)轉(zhuǎn)基因植物分析該基因的功能提供了理論依據(jù)。
[Abstract]:DNA J protein is a widespread molecular chaperone in plants, which plays an important role in maintaining the homeostasis of proteins under normal and stress conditions. By transcriptome analysis of the strong resistant plant, Aconitum mongolica, we found that a cold-drought induced J protein called Am J20 coding sequence was found in the early stage, but its 3 'terminal coding region was incomplete. In this study, the full-length sequence of c DNA was obtained by 3'RACE (800 bp), conjectured that the encoding protein consisted of 179 amino acid residues, containing a typical J domain, belonging to type 鈪,

本文編號(hào):2249004

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