本文選題：雷公藤　切入點(diǎn)：貝殼杉烯酸氧化酶(KAO)　出處：《中國中藥雜志》2017年01期

【摘要】：貝殼杉烯酸氧化酶(kaurenoic acid oxidase)是二萜赤霉素生物合成途徑上的關(guān)鍵酶,參與植物生長發(fā)育等重要生物學(xué)過程。該文根據(jù)雷公藤轉(zhuǎn)錄組數(shù)據(jù)設(shè)計(jì)特異性引物,采用PCR技術(shù)克隆得到貝殼杉烯酸氧化酶全長c DNA序列,并進(jìn)行生物信息學(xué)分析;使用實(shí)時(shí)定量PCR(qRT-PCR)研究基因的誘導(dǎo)表達(dá)水平。克隆得到Tw KAO長度為1 874 bp,編碼487個(gè)氨基酸,蛋白相對分子質(zhì)量56.02 k Da,理論等電點(diǎn)8.89;經(jīng)茉莉酸甲酯(Me JA)誘導(dǎo)后,Tw KAO基因相對表達(dá)量在12 h達(dá)到峰值;經(jīng)植株組織表達(dá)分析證實(shí),Tw KAO基因在雷公藤的葉中表達(dá)量最高,根中最低。該研究首次克隆得到雷公藤KAO基因,并分析其mRNA表達(dá)特征,為深入研究雷公藤生長發(fā)育以及萜類活性成分次生代謝研究奠定基礎(chǔ)。
[Abstract]:Kaurenoic acid oxidase is a key enzyme in the biosynthesis pathway of diterpenoid gibberellin, and is involved in plant growth and development. In this paper, specific primers were designed based on the transcriptional data of Tripterygium wilfordii. The full-length c DNA sequence of kapenoate oxidase was cloned by PCR technique and analyzed by bioinformatics. The induced expression level of the gene was studied by real-time quantitative PCR qRT-PCR.The length of Tw KAO was 1 874 BP, encoding 487 amino acids. The relative molecular weight of the protein was 56.02 kDa, the theoretical isoelectric point was 8.89, and the relative expression of Tw KAO gene reached its peak at 12 h after induction by methyl jasmonate (Me JA), and the highest expression of Tw KAO gene in the leaves of Tripterygium wilfordii was confirmed by plant tissue analysis. The KAO gene of Tripterygium wilfordii was cloned for the first time and its mRNA expression characteristics were analyzed, which laid a foundation for further study on the growth and development of Tripterygium wilfordii and the secondary metabolism of terpene active components.
【作者單位】： 首都醫(yī)科大學(xué)中醫(yī)藥學(xué)院;中國中醫(yī)科學(xué)院中藥資源中心道地藥材國家重點(diǎn)實(shí)驗(yàn)室培育基地;中醫(yī)病絡(luò)研究北京市重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金優(yōu)秀青年基金項(xiàng)目(81422053);國家自然科學(xué)基金面上項(xiàng)目(81373906) 國家杰出青年科學(xué)基金項(xiàng)目(81325023)
【分類號(hào)】：S567.19

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本文編號(hào)：1677494