菌物学报2025，Vol.44Issue(2)：68-80,13.DOI:10.13346/j.mycosystema.240172
糙皮侧耳谷氨酰胺合成酶PoGlnA的鉴定及氮源响应分析
Identification and nitrogen response of glutamine synthetase PoGlnA from Pleurotus ostreatus
摘要
Abstract
Glutamine synthetase is the key enzyme of nitrogen assimilation and plays a critical role in nitrogen metabolism.In this study,the glutamine synthetase coding gene PoGlnA belonging to the GlnA family was cloned from Pleurotus ostreatus.The PoGlnA protein was heterologously expressed and purified and the glutamine synthetase activity was determined.The results showed that the CDS sequence of PoGlnA gene was 1 446 bp,coding 481 aa.The molecular weight of PoGlnA protein was 53 kDa,possessing the glutamine synthetase activity.The phylogenetic relationship between PoGlnA protein and other reported glutamine synthetase proteins was analyzed.The result showed that there was a relatively close relationship between PoGlnA and GlnA protein from bacteria,but the relationships between PoGlnA protein and the reported glutamine synthetase protein from edible fungi were distant.The responses of PoGlnA gene to nitrogen starvation and different nitrogen sources were analyzed at the transcriptional and the translational level.The results showed that nitrogen starvation,and Asp,Asn,Leu,Glu treatment could promote PoGlnA gene expression,while ammonium and Gln treatment had no significant influence on the expression,but nitrate treatment inhibited the expression.In addition,the peptide containing dominant epitopes of PoGlnA was obtained by antigen epitope analysis,and the specific anti-PoGlnA PcAb was furtherly prepared,laying a foundation for elucidating the physiological function of PoGlnA gene.关键词
糙皮侧耳/谷氨酰胺合成酶/原核表达/酶活性/氮源响应Key words
Pleurotus ostreatus/glutamine synthetase/prokaryotic expression/enzyme activity/nitrogen response引用本文复制引用
魏志霞,刘元栋,李佳涛,胡延如,申进文,戚元成,王风芹,文晴..糙皮侧耳谷氨酰胺合成酶PoGlnA的鉴定及氮源响应分析[J].菌物学报,2025,44(2):68-80,13.基金项目
国家自然科学基金(31701980) （31701980）
国家现代农业技术体系建设专项(CARS-20) （CARS-20）
河南省科技攻关项目(232102111111) （232102111111）
河南省农业良种联合攻关项目(2022030101)This work was supported by the National Natural Science Foundation of China(31701980),the China Agriculture Research System of MOF and MARA(CARS-20),the Key Program for Science and Technology Development of Henan Province(232102111111),and the Joint Research Project for Improved Agricultural Variety in Henan Province(2022030101). （2022030101）
