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EFFECT OF ESCHERICHIA COLI IHF MUTATIONS ON PLASMID P15A COPY NUMBER

机译:大肠埃希氏菌突变对质粒P15A拷贝数的影响

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摘要

Four isogenic strains (himAhimD double mutant, himA and himD single mutants, and their wild type counterpart) harboring orip15A plasmid (pACYC184 or pACYC184Amp or pACYC177) show different copy numbers of that plasmid in the early stationary phase of cultivation. The copy number of orip15A plasmid increases about four times in the himAhimD double (65-70 copies per sell) and himD single mutant cells (50-56 copies per cell) and was almost the same in himA mutant (17-18 copies per cell) and wild type cells (14-16 copies per cell). The results suggest that HimD can form homodimers, which ore functionally competent for the regulation of orip15A plasmid copy number. Complementation experiments of himAhimD double mutant cells using plasmid carrying himA and himD genes (pP(L)hiphimA-5) confirm the effect of integration host factor (IHF) absence on increasing the copy number of orip15A plasmid (plasmid producing IHF complemented the defect of IHF mutant). The absence of IHF (using himAhimD double mutant as host) had no effect on the copy number of the pBR322 (oripMB1) plasmid. (C) 1997 Academic Press. [References: 24]
机译:带有orip15A质粒(pACYC184或pACYC184Amp或pACYC177)的四种同基因菌株(himAhimD双突变体,himA和himD单突变体,以及它们的野生型对应物)在培养的早期固定阶段显示该质粒的拷贝数不同。 orip15A质粒的拷贝数在himAhimD double(每次出售65-70拷贝)和himD单突变细胞(每个细胞50-56拷贝)中大约增加了四倍,而在himA突变体中(每个细胞17-18拷贝)几乎相同)和野生型细胞(每个细胞14-16份)。结果表明,HimD可以形成同源二聚体,其功能上足以调节orip15A质粒拷贝数。使用携带himA和himD基因的质粒(pP(L)hiphimA-5)对himAhimD双突变细胞进行的补充实验证实了整合宿主因子(IHF)缺失对orip15A质粒拷贝数增加的影响(产生IHF的质粒弥补了该缺陷) IHF突变体)。没有IHF(使用himAhimD双突变体作为宿主)对pBR322(oripMB1)质粒的拷贝数没有影响。 (C)1997学术出版社。 [参考:24]

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