Cloning and Prokaryotic Expression of Farnesyl Pyrophosphate Synthase Gene from Lepidium apetalum
MA Ligang
ZHAO Le
FU Xiaodie
FENG Weisheng
KUANG Haixue
ZHENG Xiaoke
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.
Keywords：Lepidium apetalumfarnesyl pyrophosphate synthasegene cloningsequence analysisprokaryotic expression
Publication Date：2017-09-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：6( 92-97 )