Skip to main content
. 2014 Dec 4;10(12):e1004853. doi: 10.1371/journal.pgen.1004853

Table 1. Bacterial strains and plasmids.

Bacteria Strain Relevant characteristics; comments Reference
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