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 |