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一种新型耐盐碱基因Dhodh的筛选与功能初步鉴定

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

黑龙江八一农垦大学学报2025,Vol.37Issue(2):82-89,8.
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

刘青朋 1王艳红 1徐蓓蓓 1张宇可 1朱曲辞 1王永杰 1韩露 1曹宁 1付若晗 1徐海军2

作者信息

  • 1. 黑龙江八一农垦大学生命科学技术学院/黑龙江省寒区环境微生物与农业废弃物资源化利用重点实验室,大庆 163319
  • 2. 黑龙江省科学院大庆分院
  • 折叠

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)

黑龙江八一农垦大学学报

1002-2090

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