The Escherichia coli strain DH42 is sensitive to high osmolarity in an alkaline medium. Using mini-Tn5 mutagenesis, construction of mutant strains by homologous recombination and subcloning of DNA fragment techniques, gene ompC was identified as the key factor that, once disrupted, caused osmosis-sensitivity of E. coli strain DH42 grown in an alkaline medium. Through P1 transduction, a mutant strain, D9 (W3110 ompC:kan), was constructed and growth comparison was performed between DH42 and D9 under different pHs and salt concentrations. The result showed that ompC was necessarily required for hyperosmotic adaptation of E. coli in the alkaline medium.
FabB和FabF是大肠杆菌(Escherichia.coli)脂肪酸合成的关键酶.生物信息学分析显示,粪肠球菌基因组中有2个与大肠杆菌fabF同源的基因:fabF1和fabF2,缺少与fabB同源的基因.用粪肠球菌(Enterococcus faecalis)V583总DNA为模板,PCR扩增fabF1和fabF2基因,以pBAD24为载体,构建了重组质粒pHW13(fabF1)和pHW14(fabF2).体内体外研究显示:fabF1基因能互补大肠杆菌fabB突变,FabF1具有β酮脂酰ACP合成酶Ⅰ(FabB)活性;fabF2能互补大肠杆菌fabF突变,FabF2具有β酮脂酰ACP合成酶Ⅱ(FabF)活性.同时发现粪肠球菌FabF2不同于大肠杆菌FabF,它还拥有微弱β酮脂酰ACP合成酶Ⅰ(FabB)活性,可使大肠杆菌fabB突变株产生少量的不饱和脂肪酸.上述结果表明,FabF类酶(FabF like enzyme)同样可以具有β酮脂酰ACP合成酶Ⅰ(FabB)活性.