Cloning And Expression Analysis Of FAE1 Gene From Pistacia Chinensis

Pistacia chinensis is an important kind of woody energy plant,and its seed oil can be used for making biodiesel.It has been reported that fatty acid elongase 1(FAE1) is aβ-ketoacyl-CoA synthase that can prolong the carbon chain of fatty acid,and FAE1 gene is a key factor in controlling lipid biosynthesis in Pistacia chinensis.Based on the plants FAE1 sequences recorded in the NCBI,the primers were designed,and an full-length FAE1 cDNA(named as PcFAE1) with 1972bp that contained a 1536 bp ORF was cloned by RT-PCR and RACE from seeds of Pistacia chinensis,and the bases composition,the restriction sites and the repeat fragments of PcFAE1 gene were analyzed.It is found that PcFAE1 gene shares a high sequence similarity with the sequences of cloned FAE1 genes from other species,suggesting that this cloned PcFAE1 gene might be FAE1.On this base,the physical and chemical characterization such as molecular weight and isoelectric point(pI) as well as the primary,secondary and tertiary structure of the protein encoded by PcFAE1 were analyzed and emulated by using the public database and reference softwares.The results showed that the PcFAE1 protein with 511 amino acids was a transmembrane protein located to cytoplasm,but without signal peptide,moreover,this protein has 57.81kD molecular weight with 9.28 of pI,and most of which secondary structure areα-helix and randon coli and which tertiary structure contains classical functional domain FAE1 CUTI RppA to similarity to a KCS family. Additionally,the evolutional tree of FAE1 protein was set up,and the PcFAE1 expression in E.coli was studied.The result showed that FAE1 from Pistacia chinensis and from Arabidopsis thaliana clustered together firstly.The results of the expression in E.coli showed that the maximal content of fusion protein was observed in the 1.0mmol/L IPTG treatment for 4h at 37℃.The above results may play an important role in the further identification and application of PcFAE1 gene.