Cloning and expression analysis of PlERG24 gene in ergosterol biosynthesis of Phellinus linteus / 中草药

ABSTRACT

Objective: To clone the gene full length of ergosterol C14 reductase (ERG24) in Phellinus linteus and analyze its bioinformatics and expression pattern. Methods: The primers of PlERG24 were designed according to the transcription sequence of P. linteus, the cDNA full-length sequence of PlERG24 was obtained by PCR, its bioinformatics was analyzed by Ex PASy and other online analysis software, and its expression pattern in mycelia of P. linteus was analyzed by real-time fluorescence quantitative PCR. Results: The full-length cDNA of PlERG24 gene was 1 412 bp, which encoding a protein of 441 amino acids with a predicted molecular weight of 49 358.61 and isoelectric point of 5.28; ERG24 protein was a hydrophobic protein without signal peptide, which was presumably located in the plasma membrane with six phosphorylation sites. Phylogenetic tree analysis indicated that amino acid sequences of ERG24 in P. linteus were genetically closely related to ERG24 in Sanghuangporus baumii. The qRT-PCR results showed that gene expression of PlERG24 reached the highest level of 6.36 at 25d during the growth cycle of mycelia in P. linteus. Conclusion The full length of PlERG24 gene was obtained, which lays a foundation for further studies on gene function and genetic regulatory mechanism of ergosterol biosynthesis.