IDENTIFICATION OF MUTANTS OF ESCHERICHIA-COLI DEFICIENT IN FORMATE DEHYDROGENASE AND NITRATE REDUCTASE ACTIVITIES USING DYE INDICATOR PLATES
IDENTIFICATION OF MUTANTS OF ESCHERICHIA-COLI DEFICIENT IN FORMATE DEHYDROGENASE AND NITRATE REDUCTASE ACTIVITIES USING DYE INDICATOR PLATES
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DOI：
10.1111/j.1574-6968.1977.tb00905.x
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发表时间：
1977-01-01
影响因子：
2.1
通讯作者：
HADDOCK, BA
中科院分区：
文献类型：
--
作者：
BEGG, YA;WHYTE, JN;HADDOCK, BA
人们对大肠杆菌的厌氧、质子转运、依赖形式的硝酸还原酶活性进行了广泛的研究，特别是在鉴定所需的各种氧化还原成分、它们在细胞膜中的功能组织以及负责它们合成的结构基因方面[1，2]。图1总结了迄今为止可获得的信息，从图1可以明显看出，仍有几个悬而未决的问题。我们已经开始对该系统的分子遗传学进行详细的分析，特别强调首先是目前所描述的各种结构基因产物的明确特征，其次是识别负责控制结构基因表达的调控基因。在这项研究中，很明显，以前描述的许多鉴定和相关突变体的遗传特征的方法要么耗时，要么不精确。在这里，我们描述了利用改进的染料指示板的两种方法的应用，这两种方法有助于快速和明确地将突变体分类为：(I)特别缺乏甲酸脱氢酶活性；(Ii)特别缺乏D-乳酸依赖的硝酸还原酶活性；(Iii)多向性地缺乏这两种活性。
The anaerobic, proton translocating, formatedependent nitrate reductase activity of Escherichia coli has been studied extensively, particularly with respect to the identification of the various redox components required, their functional organization in the cytoplasmic membrane, and the structural genes responsible for their synthesis [1, 2]. The information so far available is summarized in Fig. 1 from which it is apparent that several outstanding problems remain. We have begun a detailed analysis of the molecular genetics of the system with particular emphasis on firstly the unambiguous characterizations of the products of the various structural genes so far described, and secondly the identification of the regulatory genes responsible for the control of structural gene expression. In this study it became apparent that many of the previously described methods for the identification and genetic characterization of relevant mutants were either time-consuming or imprecise. We describe here the application of two methods utilizing modified dye indicator plates which facilitate the rapid and unambiguous classification of mutants into those which:(i) specifically lack formate dehydrogenase activity;(ii) specifically lack D-lactate-dependent nitrate reductase activity; and (iii) pleiotropically lack both activities.