本文選題：獨(dú)行菜 + 法尼基焦磷酸合酶�。� 參考：《河南農(nóng)業(yè)科學(xué)》2017年09期

【摘要】：從獨(dú)行菜(Lepidium apetalum)中克隆強(qiáng)心苷合成途徑的法尼基焦磷酸合酶(FPS)基因,并在大腸桿菌中表達(dá),為進(jìn)一步研究獨(dú)行菜強(qiáng)心苷的合成途徑提供參考。分析前期獲得的獨(dú)行菜幼苗轉(zhuǎn)錄組數(shù)據(jù),選擇其中注釋為FPS且具有完整開(kāi)放閱讀框的序列,從獨(dú)行菜葉片c DNA中通過(guò)PCR克隆該序列,并進(jìn)行序列分析。結(jié)果表明,克隆得到獨(dú)行菜FPS基因的cDNA序列,Gen Bank登錄號(hào)KY366218,命名為L(zhǎng)a FPS。序列長(zhǎng)度為1 332 bp,含有1 161 bp的開(kāi)放閱讀框,推測(cè)編碼386個(gè)氨基酸。La FPS蛋白可能不含跨膜結(jié)構(gòu),定位于線粒體中,含有聚異戊二烯合成酶結(jié)構(gòu)域。La FPS蛋白氨基酸序列與擬南芥(Arabidopsis thaliana)FPS1蛋白相似性高達(dá)92%,進(jìn)化樹(shù)分析結(jié)果表明,La FPS與同為十字花科的擬南芥FPS1、遏藍(lán)菜(Noccaea caerulescens)FPS1、高山南芥(Arabis alpina)FPS親緣關(guān)系最近。構(gòu)建的載體p ET-32a-La FPS可在大腸桿菌中成功誘導(dǎo)表達(dá)。表明成功克隆了獨(dú)行菜FPS基因,并建立了其原核表達(dá)體系。
[Abstract]:The FPS gene of cardiac glycoside synthesis pathway was cloned from Lepidium apetalumin and expressed in E. coli, which provides a reference for further study on the synthesis pathway of cardiac glycoside. The transcriptome data obtained in the early stage were analyzed, and the sequence annotated as FPS with complete open reading frame was selected. The sequence was cloned by PCR from the leaves of C. DNA, and the sequence was analyzed. The results showed that the cDNA sequence of FPS gene was cloned into GenGen Bank accession number KY366218, named La FPS. The sequence length was 1 332 BP, containing an open reading frame of 1 161 BP, which suggested that the 386-amino acid .La FPS protein might not contain transmembrane structure and was located in mitochondria. The amino acid sequence of the polyisoprene synthase domain. La FPS protein was similar to Arabidopsis thaliana)FPS1 protein of Arabidopsis. The phylogenetic tree analysis showed that La FPS was closely related to FPS1 of Arabidopsis, Noccaea caerulescensus FPS1 and Arabis alpina)FPS of Alpine. The constructed vector p ET-32a-La FPS can be successfully induced in E. coli. The results showed that the FPS gene was cloned successfully and its prokaryotic expression system was established.
【作者單位】： 河南中醫(yī)藥大學(xué)藥學(xué)院;呼吸疾病診療與新藥研發(fā)河南省協(xié)同創(chuàng)新中心;黑龍江中醫(yī)藥大學(xué)藥學(xué)院;
【基金】：國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展(973)計(jì)劃項(xiàng)目(2013CB531802) 河南中醫(yī)學(xué)院博士科研基金項(xiàng)目(BSJJ2011-18)
【分類號(hào)】：S567.2
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本文編號(hào)：1778900