河南农业科学2017，Vol.46Issue(9)：92-97,6.DOI:10.15933/j.cnki.1004-3268.2017.09.017
独行菜法尼基焦磷酸合酶基因的克隆与原核表达
Cloning and Prokaryotic Expression of Farnesyl Pyrophosphate Synthase Gene from Lepidium apetalum
摘要
Abstract
This study obtained the farnesyl pyrophosphate synthase gene involved in the cardiac glycosides biosynthesis from Lepidium apetalum,analysed the sequence,and induced expression of the gene in E.coli,which provided new material for research of the biosynthetic pathway of cardiac glycoside in L.apetalum.Specific primers were designed for a gene fragment with complete ORF and annotated as FPS in the transcriptome data of L.apetalum,and the fragment was cloned from L.apetalum leaf cDNA template by PCR method.Then the sequence of cloned gene was analyzed.The cloned cDNA fragment of LaFPS(GenBank accession No.KY366218) had an length of 1 332 bp,with an ORF of 1 161 bp encoding 386 amino acids.According to the sequence analysis result,LaFPS had no transmembrane helices,located in mitochondria,and had an polyprenyl synthetase domain.Sequence of LaFPS was 92％ identical to that of Arabidopsis FPS1 protein.LaFPS was closest to Arabidopsis FPS1,Noccaea caerulescens FPS1,Arabis alpine FPS in the phylogeny tree analysis,and all the proteins were from the Brassicaceae family.Expression of LaFPS protein was successfully induced in E.coli strain BL21(DE3) with constructed expression vector pET-32a-LaFPS.LaFPS gene was cloned from L.apetalum,and the prokaryotic expression system was established.关键词
独行菜/法尼基焦磷酸合酶/基因克隆/序列分析/原核表达Key words
Lepidium apetalum/farnesyl pyrophosphate synthase/gene cloning/sequence analysis/prokaryotic expression分类
农业科技引用本文复制引用
马利刚,赵乐,付小蝶,冯卫生,匡海学,郑晓珂..独行菜法尼基焦磷酸合酶基因的克隆与原核表达[J].河南农业科学,2017,46(9):92-97,6.基金项目
国家重点基础研究发展(973)计划项目(2013CB531802) （973）
河南中医学院博士科研基金项目(BSJJ2011-18) （BSJJ2011-18）
