兰州大学学报（医学版）2024，Vol.50Issue(2)：12-17,6.DOI:10.13885/j.issn.1000-2812.2024.02.003
核酸酶NucB高效表达工程菌株的构建及功能测定
Construction and functional evaluation of an engineered bacte-rial strain for high-efficiency expression of NucB nuclease
摘要
Abstract
Objective To objective of this study was to construct an engineered bacterial strain capable of effi-ciently and stably expressing NucB nuclease.Methods The DH5α(DE3)strain of Escherichia coli was used as the host organism.Gene knockout was performed using genetic editing techniques to disrupt the essential gene lexA within the bacterial genome.Subsequently,a modified plasmid containing the lexA gene was designed and introduced to complement the knockout,resulting in the engineered strain DH5α(DE3)ΔlexA.A plasmid carrying the target gene nucB was then transformed into the modified strain for fermentation cultivation and production of NucB nuclease,followed by functional characterization.Results A comparative analysis between the genetically modified strain DH5α(DE3)ΔlexA and the wild-type Escherichia coli revealed that the modi-fied strain exhibited highly stable plasmids,significantly increased protein expression levels,and the ability to produce NucB nuclease during fermentation without the need for antibiotic supplementation.Conclusion In summary,we successfully constructed an engineered strain by using genetic editing methods,DH5α(DE3)ΔlexA,which enables antibiotic-free,efficient,and stable expression of NucB nuclease.关键词
基因敲除/无抗生素/高效稳定/核酸酶NucBKey words
gene knockout/antibiotic-free/efficient and stable/NucB nuclease分类
生物科学引用本文复制引用
李江勇,陈熙明,刘光琇,张威,陈拓,袁亚玲,李善家..核酸酶NucB高效表达工程菌株的构建及功能测定[J].兰州大学学报（医学版）,2024,50(2):12-17,6.基金项目
甘肃省科技重大专项资助项目(22ZD6WA035) （22ZD6WA035）
