香菇LeRPS15A蛋白CDS序列和启动子区的克隆、生物信息学分析及干扰表达工程菌株的构建OACHSSCD

In order to construct the interference expression vector for the LeRPS15A protein gene of the mushroom ribosomal small subunit,the strain 9608 was used as material.The CDS sequence of the RPS15A protein and its promoter region were respectively cloned.Through bioinformatics analysis,the protein characteristics and evolu-tionary status of the protein were predicted.The interference expression vector was constructed to provide a basis for further functional research.The results showed that the full-length CDS sequence of the RPS15A protein in the mushroom was 393 bp,encoding 130 amino acids.The predicted molecular weight of the protein was 14.6 kDa,the isoelectric point was 10.33,the instability index was 40.89,and the average hydrophilicity was-0.005,making it an unstable hydrophilic protein.Based on the phylogenetic analysis of the amino acid se-quence,its closest relationship was with the Coprinopsis cinerea and Agaricus bisporus.The promoter region con-tained various light response,stress response and hormone response elements.Based on the cloned sequence,the interference expression vector GpiE-RPS15A integrating the promoter region and the CDS sequence was suc-cessfully constructed,and the engineered Agrobacterium rhizogenes EHA105 strain was obtained.To further elu-cidate the potential critical role of the LeRPS15A protein from Lentinula edodes in the processes of detoxification and yield enhancement of Lentinula edodes,and to provide essential experimental materials for subsequent re-search.