發(fā)布時(shí)間：2018-03-03 08:45

  本文選題：太子參　切入點(diǎn)：三萜皂苷　出處：《中草藥》2017年13期 　論文類型：期刊論文

【摘要】：目的對(duì)太子參三萜類皂苷生物合成關(guān)鍵調(diào)控酶鯊烯環(huán)氧酶1(SQE1)基因進(jìn)行全長(zhǎng)c DNA克隆和功能分析。方法基于其他植物SQE基因的同源序列設(shè)計(jì)簡(jiǎn)并引物,以太子參塊根總RNA為模板,RT-PCR結(jié)合RACE技術(shù)克隆太子參SQE1基因的全長(zhǎng)c DNA,并進(jìn)行生物信息學(xué)分析。利用農(nóng)桿菌葉盤轉(zhuǎn)化法將SQE1基因轉(zhuǎn)化煙草,研究其對(duì)煙草總?cè)�萜量的影響。結(jié)果獲得太子參SQE1基因全長(zhǎng)c DNA序列2 038 bp,該序列包含1 554 bp的開放閱讀框,編碼517個(gè)氨基酸,相對(duì)分子質(zhì)量為5.67×104,等電點(diǎn)為8.8,與其他藥用植物的SQE蛋白具有較高的同源性,含有FAD結(jié)合結(jié)構(gòu)域和4個(gè)跨膜區(qū)域,轉(zhuǎn)太子參SQE1基因的煙草的總?cè)�萜量明顯高于非轉(zhuǎn)基因植株。結(jié)論首次克隆獲得太子參SQE1基因的全長(zhǎng)c DNA,該基因的異源表達(dá)可一定程度提高轉(zhuǎn)基因植物的總?cè)�萜�?為闡明與應(yīng)用太子參三萜類成分生物合成途徑提供科學(xué)依據(jù)。
[Abstract]:Objective to clone and analyze the full-length c DNA of squalene cyclooxygenase 1 (squalene cyclooxygenase 1) gene, a key regulator of triterpenoid saponins biosynthesis of Pseudostellaria heterophylla. Methods degenerate primers were designed based on the homologous sequence of SQE gene in other plants. The full-length cDNA of SQE1 gene of Pseudostellariae heterophylla was cloned by RT-PCR and RACE using total root RNA of P. princeliae as template. Bioinformatics analysis was carried out. The SQE1 gene was transformed into tobacco by Agrobacterium tumefaciens transformation method. Results the full-length c DNA sequence of Radix Pseudostellariae SQE1 gene was 2 038 BP, which contained an open reading frame of 1 554 BP, encoding 517 amino acids. The relative molecular weight is 5.67 脳 10 ~ 4 and the isoelectric point is 8.8. It has high homology with other medicinal plants' SQE protein, and contains FAD binding domain and four transmembrane domains. Conclusion the total triterpenoids of the transgenic plants were significantly higher than that of the non-transgenic plants. Conclusion the full-length c-DNA of the SQE1 gene was obtained for the first time, and the heterologous expression of the gene could improve the total triterpenoid quantity of the transgenic plants to some extent. It provides scientific basis for elucidation and application of biosynthesis pathway of triterpenoids of Pseudostellaria heterophylla.
【作者單位】： 福建農(nóng)林大學(xué)生命科學(xué)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(30900915) 福建省教育廳資助高校重點(diǎn)項(xiàng)目(JK2012012)
【分類號(hào)】：S567.53

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