本文選題：獨行菜 + 法尼基焦磷酸合酶��； 參考：《河南農業(yè)科學》2017年09期

【摘要】：從獨行菜(Lepidium apetalum)中克隆強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,為進一步研究獨行菜強心苷的合成途徑提供參考。分析前期獲得的獨行菜幼苗轉錄組數據,選擇其中注釋為FPS且具有完整開放閱讀框的序列,從獨行菜葉片c DNA中通過PCR克隆該序列,并進行序列分析。結果表明,克隆得到獨行菜FPS基因的cDNA序列,Gen Bank登錄號KY366218,命名為La FPS。序列長度為1 332 bp,含有1 161 bp的開放閱讀框,推測編碼386個氨基酸。La FPS蛋白可能不含跨膜結構,定位于線粒體中,含有聚異戊二烯合成酶結構域。La FPS蛋白氨基酸序列與擬南芥(Arabidopsis thaliana)FPS1蛋白相似性高達92%,進化樹分析結果表明,La FPS與同為十字花科的擬南芥FPS1、遏藍菜(Noccaea caerulescens)FPS1、高山南芥(Arabis alpina)FPS親緣關系最近。構建的載體p ET-32a-La FPS可在大腸桿菌中成功誘導表達。表明成功克隆了獨行菜FPS基因,并建立了其原核表達體系。
[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)藥大學藥學院;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)藥大學藥學院;
【基金】：國家重點基礎研究發(fā)展(973)計劃項目(2013CB531802) 河南中醫(yī)學院博士科研基金項目(BSJJ2011-18)
【分類號】：S567.2
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本文編號：1778900