TABLE 2.
Plasmids used in this study
| Plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| pGEM-TEasy | PCR cloning vector; Ampr | Promega |
| pVA838 | E. coli-Streptococcus shuttle vector; Tetr Ermr | 17 |
| GFP-pVPT | Modified pVA838 with maltose promoter; Tetr | 4 |
| pAL60 | gtf2 with flanking regions in pGEM-T Easy; Ampr | This study |
| pAL61 | gtf2 gene deleted from pAL60; Ampr | This study |
| pAL62 | aphA3 gene inserted into pAL61; Ampr Kanr | This study |
| pAL63 | gtf2::aphA3 fragment cloned into pSF143; Kanr | This study |
| pGEX6p1 | Vector used to generate GST fusion proteins; Ampr | Amersham |
| pAL70 | gtf1 gtf2 fragment with extra 600-bp up- and downstream sequences in pGEM-T Easy vector; Ampr | This study |
| pAL71 | Large portion of gtf1 gtf2 fragment deleted from pAL70 by inverse PCR and self-ligated; Ampr | This study |
| pAL72 | aphA3 gene inserted in pAL71; Ampr | This study |
| pAL73 | gtf1 gtf2::aphA3 fragment cloned into pSF143; Kanr Tetr | This study |
| pVT1175 | Full-length fap1 in pHSG576; Kanr | 37 |
| pAL80 | RII deleted from pVT1175 with inverse PCR; Kanr | This study |
| pAL81 | HindIII fragment of pVA838 with an Ermr cassette and streptococcal replicon ligated with pAL80; Ermr | This study |
| pAL90 | gtf1 cloned in GFP-pVPT; Ermr | This study |
| pAL91 | gtf2 cloned in GFP-pVPT; Ermr | This study |
| pAL92 | gtf1 gtf2 fragment cloned in GFP-pVPT; Ermr | This study |
| pAL200 | gtf1 gtf2 ORFs cloned in pGEX6p1; Ampr | This study |
| pAL201 | gtf1 aa 21∼485 deleted from pAL200; Ampr | This study |
| pAL202 | gtf2 aa 15∼417 deleted from pAL200; Ampr | This study |
| pAL94 | gtf2 gene cloned into pET27b; Kanr | This study |
| pAL205 | T7 tag fused at C-terminal gtf2 in pAL200; Ampr | This study |
| pAL206 | T7 tag fused at C-terminal gtf2 in pAL201; Ampr | This study |
| pAL207 | Tn5 transposon inserted in Gtf1 aa 31 in pAL205; Ampr | This study |
| pAL208 | Tn5 transposon inserted in Gtf1 aa 65 in pAL205; Ampr | This study |
| pAL209 | Tn5 transposon inserted in Gtf1 aa 159 in pAL205; Ampr | This study |
| pAL300 | Tn5 transposon inserted in Gtf1 aa 416 in pAL205; Ampr | This study |