食用菌学报2026，Vol.33Issue(2)：32-40,9.DOI:10.16488/j.cnki.1005-9873.2026.02.003
梯棱羊肚菌谷氨酸脱氢酶基因克隆及原核表达
Cloning and Prokaryotic Expression of Glutamate Dehydrogenase Gene from Morchella importuna
摘要
Abstract
The glutamate dehydrogenase gene from Morchella importuna(MiGDH)was cloned using RT-PCR and subjected to bioinformatics analyses.A prokaryotic expression vector for MiGDH,pEB-MiGDH,was constructed and then induced for heterologous expression in Escherichia coli BL21(DE3).The results showed that the cDNA sequence of MiGDH contains an open reading frame of 1 377 bp,encoding a protein of 458 amino acids.The full-length genomic DNA of MiGDH is 1 529 bp,comprising three exons and two introns.The deduced MiGDH is a stable hydrophilic protein,and has a relative molecular mass of 50 167.51 and a theoretical isoelectric point(pI)of 5.91.It lacks a signal peptide and transmembrane domains,indicating that it is localized in the cytoplasm.The N-terminus of MiGDH contains an ELFV_dehydrogenase domain,which is identified as an NADP(H)-specific glutamate dehydrogenase.The secondary structure of MiGDH primarily consists of α-helix and random coils,with a tertiary structure forming a homologous hexamer.MiGDH was successfully expressed in the prokaryotic expression system.The heterologous expression of MiGDH provided a reference for further investigations into the biological functions of MiGDH in M.importuna.关键词
梯棱羊肚菌/谷氨酸脱氢酶/基因克隆/原核表达Key words
Morchella importuna/glutamate dehydrogenase/gene cloning/prokaryotic expression引用本文复制引用
付亚娟,张慧敏,庞盈,乔洁,侯晓强..梯棱羊肚菌谷氨酸脱氢酶基因克隆及原核表达[J].食用菌学报,2026,33(2):32-40,9.基金项目
河北省高等学校科学技术研究项目(ZD2022107) （ZD2022107）
河北省自然科学基金资助项目(H2020408003) （H2020408003）
