Table 2.
Descriptions and sources of plasmids used in this study
| Plasmids | Description | Sources |
|---|---|---|
| pTrc99a | AmpR, pBR322-origin, trc promoter, 4.2 kb | [35, 38] |
| pCas | KanR, Cas9 nuclease expression plasmid, temperature-sensitive origin | [39, 40] |
| pTrc-ilvA* | AmpR, ilvA* cloned in the sac I and BamH I site of pTrc99A | This study |
| Gap-ilvA* | AmpR, replacement of Trc promoter in pTrc-ilvA* with the Gap promoter | This study |
| Pbs-ilvA* | AmpR, replacement of Trc promoter in pTrc-ilvA* with the Pbs promoter | This study |
| pTrc-ilvA* -BleuDH | AmpR, BleuDH cloned in the Xba I and Hind III site of pTrc-ilvA* | This study |
| pTrc-ilvA*-leuDH | AmpR, leuDH cloned in the Xba I and Hind III site of pTrc-ilvA* | This study |
| pTrc-ilvA*-Trc-leuDH | AmpR, leuDH carrying a Trc promoter and RBS cloned in the Xba I and Hind III site of pTrc-ilvA* | This study |
| pTrc-ilvA*-Gap-leuDH | AmpR, leuDH carrying a Gap promoter and RBS cloned in the Xba I and Hind III site of pTrc-ilvA* | This study |
| pTrc-ilvA*-Pbs-leuDH | AmpR, leuDH carrying a Pbs promoter and RBS cloned in the Xba I and Hind III site of pTrc-ilvA* | This study |
| Gap-ilvA*-leuDH | AmpR, replacement of Trc promoter in pTrc-ilvA*-leuDH with the Gap promoter | This study |
| Gap-ilvA*-Trc-leuDH | AmpR, leuDH carrying a Trc promoter and RBS cloned in the Xba I and Hind III site of Gap-ilvA* | This study |
| Gap-ilvA*-Gap-leuDH | AmpR, leuDH carrying a Gap promoter and RBS cloned in the Xba I and Hind III site of Gap-ilvA* | This study |
| Gap-ilvA*-Pbs-leuDH | AmpR, leuDH carrying a Pbs promoter and RBS cloned in the Xba I and Hind III site of Gap-ilvA* | This study |
| Pbs-ilvA*-leuDH | AmpR, replacement of Trc promoter in pTrc-ilvA*-leuDH with the Pbs promoter | This study |
| Pbs-ilvA*-Trc-leuDH | AmpR, leuDH carrying a Trc promoter and RBS cloned in the Xba I and Hind III site of Pbs-ilvA* | This study |
| Pbs-ilvA*-Gap-leuDH | AmpR, leuDH carrying a Gap promoter and RBS cloned in the Xba I and Hind III site of Pbs-ilvA* | This study |
| Pbs-ilvA*-Pbs-leuDH | AmpR, leuDH carrying a Pbs promoter and RBS cloned in the Xba I and Hind III site of pbs-ilvA* | This study |
| pTarget-X | A plasmid used to transcript sgRNA targeting the particular gene X in genome. X refers to the amino acid biosynthetic pathways | This study |
Sp spectinomycin, Kan kanamycin, Amp ampicillin, R resistance