内蒙古民族大学学报（自然科学版）2025，Vol.40Issue(4)：10-15,6.DOI:10.14045/j.cnki.15-1220.2025.04.002
过表达yfiA、rmf和hpf对大肠杆菌DNA复制起始的影响
The Influence of Overexpression of yfiA,rmf and hpf on the Initiation of DNA Replication in Escherichia coli
摘要
Abstract
To explore the effects of overexpression of yfiA,rmf and hpf on the initiation of DNA replication in Escherichia coli by detecting the cell phenotypic changes after overexpression of the corresponding genes in the ribo-somal hibernation factor yfiA,rmf and hpf deletion mutations(ΔyfiA,Δrmf and Δhpf).The recombinant plasmid pA-CYC177-yfiA,pACYC177-rmf and pACYC177-hpf were constructed using the moderate copy plasmid pACYC177,and they were transferred into the ΔyfiA,Δrmf and Δhpf mutants respectively,the empty plasmid pACYC177 was used as the control.The phenotypic changes of bacteria after overexpression of the missing gene were observed,in-cluding DNA replication patterns,cell doubling time,cell size,changes in total intracellular protein and DnaA pro-tein content.The results showed that compared with the control BW25113/pACYC177,after overexpression of the corresponding genes in ΔyfiA,Δrmf and Δhpf mutants,there was a mild premature occurrence of DNA replication initiation,the cell doubling time was the same or shortened,the cell size still decreased,and the relative amount of intracellular total protein was 95%,90%and 92%,respectively.The relative amount of DnaA protein was 98%,93%and 110%.These results indicated after overexpression of the missing gene in the mutants,the DNA replica-tion pattern,cell doubling time,intracellular total protein and DnaA protein content restored towards to the wild type,but the cell size did not restore towards to the wild type.The possible reason that the total intracellular protein was still lower than that of the wild type after overexpression of the missing gene.关键词
yfiA/rmf/hpf/pACYC177质粒/DNA复制起始/细胞表型Key words
yfiA/rmf/hpf/plasmid pACYC177/DNA replication initiation/cell phenotype分类
生物科学引用本文复制引用
张树军..过表达yfiA、rmf和hpf对大肠杆菌DNA复制起始的影响[J].内蒙古民族大学学报（自然科学版）,2025,40(4):10-15,6.基金项目
内蒙古自治区自然科学基金项目(2019LH03017) （2019LH03017）
内蒙古自治区直属高校基本科研业务费项目(GXKY23Z055) （GXKY23Z055）
内蒙古民族大学博士科研启动基金项目(BS530) （BS530）
