本文關(guān)鍵詞： 黃花草木樨 MoSOS基因 同源克隆 序列分析 基因表達(dá)　出處：《生物技術(shù)通報(bào)》2017年09期 　論文類型：期刊論文

【摘要】：植物質(zhì)膜Na+/H+逆向轉(zhuǎn)運(yùn)蛋白基因SOS1是植物耐鹽性必需的基因之一,在抵御鹽脅迫過程中發(fā)揮十分重要的作用。以黃花草木樨葉片總RNA為模板,通過RT-PCR結(jié)合RACE方法克隆得到黃花草木樨MoSOS1基因全長(zhǎng)序列,命名為MoSOS1。序列分析表明該基因全長(zhǎng)為3 931 bp,開放閱讀框(ORF)為2 874 bp,編碼957個(gè)氨基酸,分子量為112.8 k D,等電點(diǎn)為5.31。TMHAM軟件跨膜區(qū)的預(yù)測(cè)分析表明,黃花草木樨MoSOS1蛋白具有8個(gè)跨膜結(jié)構(gòu)區(qū)域,N端和C端都位于細(xì)胞外。氨基酸序列分析表明,MoSOS1蛋白含有1個(gè)Na+/H+Exchanger superfamily和一個(gè)c NMP(Cyclic nucleotide-monophosphate)結(jié)合位點(diǎn)以及1個(gè)CAP_ED(Catabolite gene activator protein-effector domain)superfamily結(jié)構(gòu)域。生物信息預(yù)測(cè)顯示,MoSOS1的編碼蛋白為不穩(wěn)定酸性蛋白,不存在信號(hào)肽,二級(jí)結(jié)構(gòu)多為α-螺旋和無規(guī)則卷曲。熒光實(shí)時(shí)定量RT-PCR分析表明:隨著Na Cl濃度的增加,黃花草木樨地上部和根中MoSOS1基因表達(dá)水平呈增加趨勢(shì),根中表達(dá)量大于地上部,表明MoSOS1基因的表達(dá)受鹽脅迫誘導(dǎo)和調(diào)節(jié)。
[Abstract]:Plant plasma membrane Na / H antiporter gene SOS1 is one of the essential genes of plant salt tolerance and plays an important role in resisting salt stress. The total RNA of sweet clover leaves was used as a template. The full-length MoSOS1 gene of Melilotus lutei was cloned by RT-PCR and RACE, and named MoSOS1.The sequence analysis showed that the full-length of the gene was 3 931 BP, and the open reading frame (ORF) was 2 874 BP, encoding 957 amino acids. The predicted molecular weight is 112.8 KD and the isoelectric point is 5.31.TMHAM. The N-terminal and C-terminal of the MoSOS1 protein are located outside the cell. Amino acid sequence analysis shows that the moosos1 protein contains a Na / H Exchanger superfamily and a c NMP(Cyclic nucleotide-monophosphate) binding site and a CAP_ED(Catabolite gene activator protein-effector domain)superfamily node. Domain. Bioinformatics prediction shows that the encoded protein of MoSOS1 is unstable acidic protein. There was no signal peptide, and the secondary structure was mostly 偽 -helix and irregular curl. Fluorescence real-time quantitative RT-PCR analysis showed that with the increase of NaCl concentration, the level of MoSOS1 gene expression in the upper part and root of Xidi showed an increasing trend. The expression of MoSOS1 gene in root was higher than that in shoot, which indicated that the expression of MoSOS1 gene was induced and regulated by salt stress.
【作者單位】： 北京市農(nóng)林科學(xué)院北京草業(yè)與環(huán)境研究發(fā)展中心;
【基金】：國(guó)家國(guó)際科技合作專項(xiàng)(2015DFR30570) 北京市農(nóng)林科學(xué)院科技創(chuàng)新能力建設(shè)專項(xiàng)(KJCX20170110)
【分類號(hào)】：Q943.2
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本文編號(hào)：1551473