Gene Cloning, Bioinformatic Analysis and Eukaryotic Expression Vector Construction of FIP-gap Gene from Ganoderma applanatum
Ganoderma applanatum is a special type of Ganoderma spieces.In this study,a novel gene was cloned from G.applanatum and analyzed through Bioinformatics,which could supply some basis for effectively express of FIP-gap and investigation its biological functions.Using Genome walking method,the noval FIP-gap gene was amplified from the genomic DNA of G.applanatum mycelia.The nucleotide sequence analysis showed that FIP-gap comprises 342 bps and encodes a protein with 113 amino acids.The protein sequence analysis of FIP-gap indicated that molecular mass of FIP-gap is 12.7 kD and pI is 4.93.There are rich in Serines,Valines and Threonines in the peptide of FIP-gap,without Histidines or Cysteines.The homology analysis based on protein sequence blasting indicated that FIP-gap contains typical conserved regions of FIPs,and shares 78％,78％,78％,79％,78％ and 77％ homology with LZ-8 (G.lucidum),FIP-gts (G.tsugae),FIP-gja (G.japoncium),FIP-gsi (G.sinensis),FIP-gmi (G.microsporum) and FIP-gat (G.atrum),respectively.The phylogenetic tree analysis showed that FIP-gap doesn't cluster with any other FIPs from Ganoderma and forms a unique line,which implies its long distance of molecular evolution with the other FIPs.Also,the recombinant expression plasmid pPIC9-FIP-gap was constructed after the FIP-gap gene was inserted in a Pichia expression vector pPIC9.Conclusively,this study will provide helpful basis to further study on the bio-functions of FIP-gap,and full understanding of the FIPs.
国家科技期刊平台
NSTL
万方数据
维普
