Hanke P D, Fuchs J A
J Bacteriol. 1983 Dec;156(3):1192-7. doi: 10.1128/jb.156.3.1192-1197.1983.
Total Escherichia coli RNA was separated by electrophoresis on methyl mercury agarose gels, transferred to diazobenzyloxymethyl-paper, and hybridized to various DNA probes containing different segments of the nrd genes to determine the organization of these genes. A 3.2-kilobase polycistronic mRNA transcript which hybridizes to both the nrdA and nrdB genes indicated that the nrdA and nrdB genes are organized in an operon. The polycistronic transcript contained the nrdA gene at the 5' end and the nrdB gene at the 3' end. The size of the polycistronic mRNA was sufficient to code for the 80,000-molecular-weight B1 protein and the 40,000-molecular-weight B2 protein. The results also indicated that the nrdA and nrdB genes are the only genes in E. coli that code for ribonucleoside diphosphate reductase. Two smaller RNA species that hybridized to nrd DNA were observed and probably overlap with the 3.2-kilobase nrd mRNA.
通过在甲基汞琼脂糖凝胶上进行电泳分离大肠杆菌的总RNA,将其转移至重氮苄氧基甲基纸上,并与含有nrd基因不同片段的各种DNA探针杂交,以确定这些基因的组织形式。一种与nrdA和nrdB基因均杂交的3.2千碱基多顺反子mRNA转录本表明,nrdA和nrdB基因以操纵子形式组织。该多顺反子转录本在5'端含有nrdA基因,在3'端含有nrdB基因。多顺反子mRNA的大小足以编码80,000分子量的B1蛋白和40,000分子量的B2蛋白。结果还表明,nrdA和nrdB基因是大肠杆菌中仅有的编码核糖核苷二磷酸还原酶的基因。观察到两种与nrd DNA杂交的较小RNA种类,它们可能与3.2千碱基的nrd mRNA重叠。