南阳师范学院学报2026，Vol.25Issue(3)：75-82,8.
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
Cloning of the CDS Sequence and Promoter Region of the LeRPS15A Protein of the Lentinula edodes,Bioinformatics Analysis and Construction of an Interference Expression Engineering Strain
摘要
Abstract
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.关键词
香菇/LeRPS15A/基因克隆/生物信息学分析/载体构建Key words
Shiitake Mushroom/LeRPS15A/gene cloning/bioinformatic analysis/vector construction分类
农业科技引用本文复制引用
王照昱,张志峰,尚增强,石战,李建波,王栓成,白明峰,李玉英,张英君..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[J].南阳师范学院学报,2026,25(3):75-82,8.基金项目
河南省科技厅2026年度河南省重点研发专项"南水北调水源区主栽香菇品种高产关键基因验证与种质提升技术研究"(261111112000) （261111112000）
河南省科技厅关于2023年度河南省重点研发推广专项"河南主栽香菇脱毒复壮技术研究"(232102110210). （232102110210）
