Cloning and Expression Analysis of ACC Gene in Pleurotus tuoliensis
The acetyl-CoA carboxylase(ACC)gene of Pleurotus tuoliensis(PtACC)was cloned,and the encoded ACC protein was analyzed by bioinformatics tools for physicochemical properties,signal peptide,transmembrane structure,subcellular localization,secondary structure,tertiary structure and conserved domains.Based on the ACC protein sequence of P.tuoliensis(PtACC)and that of other 16 species,such as Coprinopsis cinerea,P.ostreatus,Lentinula edodes,and P.eryngii,a phylogenetic tree was constructed using the neighbor-joining algorithm.The relative expression of PtACC gene under different pH(3,4,5,6,7,8,9,10,11,and 12)was measured by fluorescence quantitative PCR.The results showed that the DNA sequence of PtACC was 7 366 bp,comprising 12 introns and 13 exons,and its cDNA was 6 696 bp,encoding a protein product of 2 231 amino acids with a molecular weight of 247.4.The PtACC protein was predicted to be an unstable acidic hydrophilic protein without any signal peptide or transmembrane structure,and is likely to present in the cytoplasm,nucleus or peroxisome.It is primarily composed of α-helix,random coil,β-sheet and β-turn.It has four conserved domains,ACC_central,AccC(BC),AccB(BCCP),and carboxyl_trans(CT).Phylogenetic analysis showed that PtACC was culstered with the ACC of P.eryngii.Compared with the control pH of 5.42,the relative expression of PtACC was significantly higher at pH 6,7,8,9,10 and 11.
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