Haworth R S, Jensen P R, Michelsen O, Wyatt J A, Brealey C J, Beechey R B
Department of Biochemistry, University College of Wales, Aberystwyth, U.K.
Biochim Biophys Acta. 1990 Aug 9;1019(1):67-72. doi: 10.1016/0005-2728(90)90125-n.
The uncoupler resistant bacterial strains E. coli Tuv and Cuv share the high deoxycholate sensitivity of the parent strain, Doc S. However, both Tuv and Cuv show greater resistance than Doc S to other detergents. Measurement of the periplasmic volume indicates that the outer membrane of Doc S is freely permeable to both TPP+ and hydroxymethylinulin. Tuv and Cuv are able to exclude these compounds. EDTA treatment was necessary prior to measuring membrane potential in Tuv and Cuv. Under conditions where delta phi could be measured, uncouplers acted to dissipate delta phi with equal potency in all strains. Uncoupler resistant proline uptake in Tuv and Cuv was abolished by EDTA treatment. Transduction experiments with phage P1 showed that uncoupler resistance could be transferred from Tuv to Doc S. Such transductants were no longer sensitive to novabiocin. The gene for uncoupler resistance cotransduced with the gene pyrE (82 min). Plating efficiency experiments with P1 suggests that detergent sensitivity in Doc S arises from an rfa (81 min) mutation. This mutation is no longer present in Tuv.
对解偶联剂具有抗性的大肠杆菌菌株Tuv和Cuv与亲本菌株Doc S一样对脱氧胆酸盐高度敏感。然而,Tuv和Cuv对其他去污剂的抗性均高于Doc S。周质体积的测量表明,Doc S的外膜对TPP⁺和羟甲基菊粉均可自由通透。Tuv和Cuv能够排除这些化合物。在测量Tuv和Cuv的膜电位之前,需要进行EDTA处理。在能够测量Δφ的条件下,解偶联剂在所有菌株中均以相同效力作用于耗散Δφ。EDTA处理消除了Tuv和Cuv中对解偶联剂具有抗性的脯氨酸摄取。用噬菌体P1进行的转导实验表明,解偶联剂抗性可从Tuv转移至Doc S。此类转导子对新生霉素不再敏感。与pyrE基因(82分钟)共转导了解偶联剂抗性基因。用P1进行的平板效率实验表明,Doc S中对去污剂的敏感性源于rfa(81分钟)突变。该突变在Tuv中不再存在。