黑龙江八一农垦大学学报2025,Vol.37Issue(2):82-89,8.DOI:10.3969/j.issn.1002-2090.2025.02.012
一种新型耐盐碱基因Dhodh的筛选与功能初步鉴定
Screening and Preliminary Functional Characterization of a Novel Salinity Tolerance Dhodh Gene
摘要
Abstract
Based on the combination of gene library and functional complementation,using pUC18 as library construction vector,the characteristic was applied that Escherichia coli KNabc could not survive in Na+environment containing 0.2 mol·L-1,the gene of improving salt tolerance of the deficient strain KNabc was successfully selected from Pseudidiomarina sediminum.Bioinformatics analysis showed that the gene encoded Dihydroorotate dehydrogenase(DHODH)and name it Ps-Dhodh.The expression vector was constructed using plasmid pET28a and Ps-Dhodh,and transferred to E.coli BL21 strain.Western blot detection of protein expression showed that the molecular weight of PS-DHODH protein was 34.03 kDa.Enzyme-linked immunosorbent assay(ELISA)showed that the enzyme activity of the overexpressed strain could reach 325 IU·L-1,which was significantly different from that of the control group.Ps-Dhodh was a novel saline-alkali tolerance gene,which belonged to a new member of DHODH protein family.The discovery enriched the saline-alkali tolerance gene pool and laid a foundation for the construction of salt-tolerant strains and their application in industry and agriculture.关键词
沉积物假海源杆菌/Dhodh/大肠杆菌/酶联免疫Key words
Pseudidiomarina sediminum/Dhodh/Escherichia coli/ELISA分类
生物科学引用本文复制引用
刘青朋,王艳红,徐蓓蓓,张宇可,朱曲辞,王永杰,韩露,曹宁,付若晗,徐海军..一种新型耐盐碱基因Dhodh的筛选与功能初步鉴定[J].黑龙江八一农垦大学学报,2025,37(2):82-89,8.基金项目
黑龙江省自然科学基金联合引导项目(LH2022C059) (LH2022C059)
黑龙江八一农垦大学"三纵"基础培育目(ZRCPY202028) (ZRCPY202028)
黑龙江省省属科研院所科研业务费项目(CZKYF2022-1-B025). (CZKYF2022-1-B025)