根瘤菌 meliloti 固氮基因在大肠杆菌小型细胞中的表达：氮酶还原酶亚基的定位。

Expression of Rhizobium meliloti genes for nitrogen fixation in Escherichia coli minicells: Mapping of the subunit of the nitrogenase reductase.

机构信息

Lehrstuhl für Genetik, Universität Bielefeld, D-4800, Bielefeld 1, F.R.G..

出版信息

Plant Mol Biol. 1982 Dec;1(4):305-20. doi: 10.1007/BF00027562.

Abstract

An EcoRI fragment of Rhizobium meliloti M2011 which shows homology to Klebsiella pneumoniae DNA carrying nifH and nifD was cloned in both orientations into the Cm gene of plasmid pACYC184 and expressed in Escherichia coli minicells. Fragment specific polypeptides of Mr 12 500, 21 000, 30 000, and 31 000 could be identified. By transposon mutagenesis it was shown that two of them (Mr 12 500 and 21 000) are fusion products with parts of the chloramphenicol acetyltransferase. The other two polypeptides are specified by one coding region which could be mapped by transposon mutagenesis. There are several reasons (homology to Klebsiella nifH, sequence data and molecular weight of the gene products) to assume that this coding region represents the R. meliloti nifH gene (gene for the subunit of the R. meliloti nitrogenase reductase, RmII).

摘要

将 Rhizobium meliloti M2011 的 EcoRI 片段克隆到质粒 pACYC184 的 Cm 基因中，并以两种方向在大肠杆菌小细胞中表达，该片段与携带 nifH 和 nifD 的 Klebsiella pneumoniae DNA 同源。可以鉴定出 Mr 12500、21000、30000 和 31000 的特定片段多肽。通过转座子诱变表明，其中两个（Mr 12500 和 21000）是与氯霉素乙酰转移酶部分融合的产物。另外两个多肽由一个编码区指定，该编码区可通过转座子诱变进行定位。有几个原因（与 Klebsiella nifH 的同源性、序列数据和基因产物的分子量）假设该编码区代表 R.meliloti nifH 基因（R.meliloti 氮酶还原酶亚基的基因，RmII）。