Zumft W G, Döhler K, Körner H
J Bacteriol. 1985 Sep;163(3):918-24. doi: 10.1128/jb.163.3.918-924.1985.
Transposon (Tn5) mutagenesis of Pseudomonas perfectomarina with the plasmid pSUP2021 [(pBR325-Mob(RP4))::Tn5] and the chromosomally integrated RP4 plasmid in Escherichia coli as the donor, produced three distinct groups of mutants that were defective in nitrous oxide respiration. One group of mutants lacked the structural protein of N2O reductase, the second synthesized a copper-free apoprotein; and a third group expressed a low level of intact enzyme. The mutants provided evidence for N2O being the immediate precursor of dinitrogen in denitrification and documented the essentiality of the copper enzyme. Synthesis of N2O reductase depended strongly on the growth conditions, with N2O-grown cells expressing the lowest level of enzyme. Regulatory responses of mutants elicited by nitrate or oxygen were unaltered when compared with wild-type behavior.
以携带质粒pSUP2021[(pBR325-Mob(RP4))::Tn5]的完美假单胞菌（Pseudomonas perfectomarina）以及大肠杆菌中染色体整合的RP4质粒作为供体进行转座子（Tn5）诱变，产生了三组不同的在氧化亚氮呼吸方面存在缺陷的突变体。一组突变体缺乏一氧化二氮还原酶的结构蛋白，第二组合成了一种无铜的脱辅基蛋白；第三组表达的完整酶水平较低。这些突变体为反硝化过程中一氧化二氮是氮气的直接前体提供了证据，并证明了铜酶的必要性。一氧化二氮还原酶的合成强烈依赖于生长条件，在以一氧化二氮为生长底物的细胞中该酶的表达水平最低。与野生型行为相比，硝酸盐或氧气引发的突变体的调节反应未发生改变。
