【摘要】：目的從羅馬洋甘菊Chamaemelum nobile中克隆萜類化合物合成關(guān)鍵酶吉馬烯A合成酶(germacrene A synthase,GAS)cDNA全長(zhǎng),并對(duì)其進(jìn)行生物信息學(xué)和組織表達(dá)分析。方法基于前期對(duì)羅馬洋甘菊轉(zhuǎn)錄組測(cè)序結(jié)果,設(shè)計(jì)特異性引物,利用反轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(RT-PCR)擴(kuò)增得到GAS的cDNA全長(zhǎng),利用生物信息學(xué)手段對(duì)其核苷酸和編碼的氨基酸序列進(jìn)行分析,并預(yù)測(cè)其編碼的蛋白質(zhì)的理化性質(zhì)和功能。通過實(shí)時(shí)定量PCR技術(shù)(qRT-PCR)檢測(cè)羅馬洋甘菊GAS基因(CnGAS)在羅馬洋甘菊不同組織的表達(dá)水平。結(jié)果擴(kuò)增得到的羅馬洋甘菊吉馬烯A合成酶基因的cDNA全長(zhǎng)為1 785 bp,命名為CnGAS(GenBank登錄號(hào)為KU589283),包含1個(gè)1 683 bp的開放閱讀框(ORF),編碼561個(gè)氨基酸,預(yù)測(cè)的相對(duì)分子質(zhì)量和等電點(diǎn)分別為6 470和5.21。CnGAS基因編碼的氨基酸序列與其他植物的GAS蛋白高度同源,且含有DDxx D保守序列。系統(tǒng)進(jìn)化樹表明CnGAS基因與菊科植物的GAS基因的親緣關(guān)系最近。qRT-PCR結(jié)果顯示CnGAS基因在羅馬洋甘菊各組織中均有表達(dá),且花中表達(dá)量最高。結(jié)論從羅馬洋甘菊中克隆得到CnGAS基因,分析了該基因在不同組織中的表達(dá)水平,為羅馬洋甘菊萜類化合物的生物合成代謝途徑的研究奠定了基礎(chǔ)。
[Abstract]:Aim to clone the full length of (germacrene A synthase,GAS cDNA, a key enzyme of terpenoid synthesis from chamomile Chamaemelum nobile, and to analyze its bioinformatics and tissue expression. Methods the full length of GAS was amplified by reverse transcription polymerase chain reaction (RT-PCR) based on the sequencing results of the previous transcripts of chamomile Roman chamomile, and the specific primers were designed to amplify the full length of cDNA by reverse transcription polymerase chain reaction (RT PCR). The nucleotide and amino acid sequences were analyzed by bioinformatics, and the physicochemical properties and functions of the encoded proteins were predicted. Real-time quantitative PCR (qRT-PCR) was used to detect the expression level of GAS gene (CnGAS) in different tissues of chamomile Roman chamomile. Results the full length of cDNA was 1 785 bp, named CnGAS (GenBank accession number (KU589283), which contained an open reading frame (ORF) (ORF), encoding 561 amino acids (ORF) of 1 683 bp. The predicted molecular weight and isoelectric point were 6470 and 6470 respectively. The amino acid sequence encoded by the 5.21.CnGAS gene was highly homologous to the GAS protein of other plants and contained the conserved DDxx D sequence. Phylogenetic tree showed that CnGAS gene was closely related to GAS gene of Compositae. QRT-PCR results showed that CnGAS gene was expressed in all tissues of chamomile, and the highest expression was found in flowers. Conclusion CnGAS gene was cloned from chamomile and its expression level in different tissues was analyzed, which laid a foundation for the study of biosynthetic pathway of chamomile terpenoids in Roman chamomile.
【作者單位】： 長(zhǎng)江大學(xué)園藝園林學(xué)院;靶向抗腫瘤藥物湖北省協(xié)同創(chuàng)新中心;荊楚理工學(xué)院化工與藥學(xué)院;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(31400603) 大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃項(xiàng)目(201207014)
【分類號(hào)】：R284

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 韓松林;李新霞;趙東升;何曉文;楊梅;耿晶;;新疆2種洋甘菊中5種化合物的微乳薄層色譜鑒別[J];時(shí)珍國(guó)醫(yī)國(guó)藥;2014年07期

2 辛祿德;李強(qiáng);王建玲;勞斐;田樹革;;微波消解-原子熒光光譜法測(cè)定洋甘菊中砷的含量[J];新疆醫(yī)科大學(xué)學(xué)報(bào);2014年07期

3 徐楊斌;王凱;冒德壽;李智宇;唐輝;楊眉;;氣相色譜-飛行時(shí)間質(zhì)譜法分析德國(guó)洋甘菊油中的揮發(fā)性成分[J];香料香精化妝品;2014年02期

4 楊彥松;潘浪勝;;洋甘菊中黃酮類成分的分離與結(jié)構(gòu)確定[J];應(yīng)用化工;2008年06期

5 蘭衛(wèi);郭玉婷;耿直;倪健;;德國(guó)洋甘菊花序中總皂苷的含量測(cè)定[J];新疆醫(yī)科大學(xué)學(xué)報(bào);2014年03期

6 韓松林;李新霞;勉強(qiáng)輝;蘭衛(wèi);劉巖;;薄層生物自顯影技術(shù)比較新疆2種洋甘菊抗氧化活性[J];中國(guó)中藥雜志;2013年02期

7 景玉霞;郭玉婷;蘭衛(wèi);;洋甘菊中6種微量元素含量的測(cè)定[J];新疆醫(yī)科大學(xué)學(xué)報(bào);2013年09期

8 袁藝;龍子江;楊俊杰;袁彩紅;;洋甘菊揮發(fā)油抗炎作用的研究[J];藥物生物技術(shù);2011年01期

9 蘭衛(wèi);郭玉婷;耿直;倪健;;洋甘菊中芹苷元-7-葡萄糖苷含量測(cè)定[J];新疆醫(yī)科大學(xué)學(xué)報(bào);2014年03期

10 ;[J];;年期

相關(guān)碩士學(xué)位論文 前2條

1 韓松林;新疆兩種洋甘菊質(zhì)量評(píng)價(jià)[D];新疆醫(yī)科大學(xué);2013年

2 朱棟梁;羅馬洋甘菊和箬竹葉中精油和黃酮的萃取、鑒定及應(yīng)用[D];江南大學(xué);2015年

，

本文編號(hào)：2436721