|
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
|
