L. lactis
|
IL1403 |
plasmid free; recipient strain, strain for all RTP assays; sequenced genome is available (GenBank: NC_002662) |
[53]
|
L. lactis
|
DM2036 |
donor of pRS01::pTRK28 |
[40]
|
E. coli
|
DH5α |
F−
endAI recAl hsdRl7 (rK− mK−) deoR supE44 thi-J gyrA96 relA
|
Gibco-BRL |
E. coli
|
TOP10 |
F−
mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu) 7697 galU galK rpsL (StrR) endA1 λ−; expression strain |
Invitrogen |
E. coli
|
TOP10F′ |
F′{lacIq, Tn10(TetR)}c mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu) 7697 galU galK rpsL (StrR) endA1 nupG; ssDNA production |
Invitrogen |
Plasmids
|
|
|
|
pLNRK-RIG |
D-C |
L. lactis/E. coli shuttle vector contains nisin-inducible promoter, nisR nisK, CamR, RIG cassette, intron donor |
[11], [40]
|
pCJK21 |
|
nisin-inducible expression vector, ErmR, SpcR
|
[41]
|
pRS01::pTRK28 |
D-pRS01 |
co-integrant of pRS01 and pTRK28, ErmR
|
[40]
|
pDL278 |
|
shuttle vector, SpcR
|
[54]
|
pBSΔltrB::tet
|
|
pBlueScript with ltrB partially replaced with tet gene, AmpR, TetR
|
Present study; pBlueScript – Agilent Technologies |
pRS01ΔltrB::tet
|
D-pRS01ΔltrB
|
pRS01::pTRK28 with ltrB replaced with tet gene, ErmR, TetR
|
Present study |
pLtrB::Ll.LtrB |
D-pLtrB |
full-length intron-containing ltrB, cloned into SpeI/SphI sites of pCJK21 |
Present study |
pLtrB(Y21A)::Ll.LtrB |
D-pLtrB(Y21A) |
Full-length intron-containing ltrB, cloned into SpeI/SphI sites of pCJK21, Y21A mutant |
Present study |
pLtrB(Y21F)::Ll.LtrB |
|
Y21F ltrB mutant |
Present study |
pLtrB(S117R)::Ll.LtrB |
|
S117R ltrB mutant |
Present study |
pLtrB(S117A)::Ll.LtrB |
|
S117A ltrB mutant |
Present study |
pLtrB(H159R)::Ll.LtrB |
|
H159R ltrB mutant |
Present study |
pLtrB(H159A)::Ll.LtrB |
|
H159A ltrB mutant |
Present study |
pLtrB(H161R)::Ll.LtrB |
|
H161R ltrB mutant |
Present study |
pLtrB(H161A)::Ll.LtrB |
|
H161A ltrB mutant |
Present study |
pCY20 |
|
full-length intronless ltrB, cloned into SpeI/SphI sites of pCJK21 |
[29]
|
pBAD33 |
|
expression vector, arabinose-inducible promoter; CamR
|
[50]
|
pLtrB-HIS6 |
|
full-length intronless ltrB, cloned into pBAD33, N-end HIS-tagged |
Present study |
pLtrB(Y21A)-HIS6 |
|
full-length intronless ltrB, cloned into pBAD33, Y21A mutant; N-end HIS-tagged |
Present study |
pONoriT1
|
|
oriT from pRS01 cloned into pGEM, AmpR
|
Present study |
pONoriT2
|
|
oriT from R388 cloned into pGEM, AmpR
|
Present study |
pONLLglnP-R |
|
fragment of glnP from L. lactis IL1403 cloned in reverse orientation into pGEM, AmpR
|
Present study |