Table 3 Bacterial strains, plasmids and primers used in this study
Strain/plasmid/primer | Relevant characteristics | Reference |
|---|---|---|
Strains | ||
NCIMB 8052 | Wild-type | NCIMB |
3304::int | Group II intron inserted at 101/102a of Cbei_3304 | This study |
3304::cp | 3304::int harboring 3304-expression vector | This study |
3304::int-pWD1 | 3304::int harboring control vector pWD1 | This study |
E. coli DH5α | Cloning | Invitrogen |
E. coli Top10 | Harboring pAN2 plasmid | Invitrogen |
Plasmids | ||
pAN2 | Methylation plasmid, Φ3T I, p15A ori, TetR | [20] |
pWJ1 | Derived from pSY6 with pCB102 ORI instead of pIM13 ORI | [21] |
pWJ1-3304 | Derived from pWJ1 for intron insertion in Cbei_3304 at 101/102a | This study |
pWD1-3304 | Derived from pWJ1, with Cbei_3304 expressing cassette added | This study |
Primers | Sequence (5′→3′) | |
pWJ1-101-F | GGAGTGTCGAGGATCCTCGAGATTATCCTTATAATTCAAAGC | This study |
pWJ1-101-R | GGTTCTCCTACAGATTGTACAAGTCAAAGCACATAACTTAC | This study |
pWD1-101-F | GGAGTGTCGAGGATCCTCGAGATGACAAAGGTTAATAAATT | This study |
pWD1-101-R | CAGATTGTACTGAGAGTGCACCATATGTAATCAACATTGATGGAAT | This study |
3304-Test-F | ATGCGGATCCATGACAAAGGTTAATAAATT | This study |
3304-Test-R | GTACGAATTCTTAATCAACATTGATGGAAT | This study |