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Table 2 Plasmids used in these studies

From: Use of nfsB, encoding nitroreductase, as a reporter gene to determine the mutational spectrum of spontaneous mutations in Neisseria gonorrhoeae

Plasmids Properties Source
pK18 General cloning vector [38]
pHP45Σ Plasmid containing the Σ interposon [39]
pNFSB The nfsB region from FA1090 was amplified by PCR using primers NP1 and NP2. The amplicon was purified, digested with BamHI and cloned into the BamHI site in pK18. This study
pEC1 The DNA between the adjacent BsmI sites were removed by digesting pEC2 with BsmI, ligating the DNA and transforming it into E. coli. This study
pEC2 Two BsmI sites were inserted into pNFSB by PCR amplification using primers NfsBBsmI-3F and -2R, treating the amplicon with S1 nuclease and polynucleotide kinase, ligating the DNA and transforming it into E. coli. This study
pEC3 A BsrGI site was introduced downstream of the NfsB coding sequence by PCR amplification of pEC1 using primers dwnstrm-F and dwnstrm-R. The amplicon was digested with BsrGI, ligated with a DNA fragment encoding the Σ interposon (amplified from pHP45Σ using Primer OmegaABC and digested with BsrGI) and transformed into E. coli. This study