Primersa
| |
This study
|
PaispS-F
|
AGACGGTCTGCCAATTATGAACC
|
This study
|
PaispS-R
|
CTCTAGATTATCTCTCAAAGGGTAGAATAG
|
This study
|
PtispS-F
|
GCC
ATG
GCATGTTCTGTAAGCACAG
|
This study
|
PtispS-R
|
CTCTAGATTATCTCTCAAAGGGTAGAATAG
|
This study
|
KispS-F
|
TCTCTGGAAAATGACCTTAAGG
|
This study
|
KispS-R
|
CTCTAGATTAGCAGCCGGATCCCACGTAC
|
This study
|
sPtispS-F
|
GCC
ATG
GCTTGCTCTGTATCCAC
|
This study
|
sPtispS-R
|
CTCTAGATTAGCGTTCGAACGGCAGAATTG
|
This study
|
MVA-F
|
GTCTAGATACCTGACGCTTTTTATCGCAAC
|
This study
|
MVA-R
|
GTCTAGAGTTTCGATAAGAGCGAACGG
|
This study
|
Plasmids
|
pTrc99A
|
Ptrc expression vector, pBR322 origin, lacIq, Ampr
|
[23]
|
pTrc99S
|
pTrc99A containing strong RBS
|
This study
|
pSTV28
|
Plac expression vector, pACYC184 origin, lacZ, Cmr
|
Takara
|
pT-PaispS
|
pTrc99A containing ispS from P. alba
|
This study
|
pT-PtispS
|
pTrc99A containing ispS from P. trichocarpa
|
This study
|
pT-KispS
|
pTrc99A containing ispS from Kudzu vine
|
This study
|
pT-sPtispS
|
pTrc99A containing codon optimized ispS from P. trichocarpa
|
This study
|
pTS-sPtispS
|
pTrc99S containing codon optimized ispS from P. trichocarpa
|
This study
|
pTS-sPt-MVA
|
pTS-sPtispS containing MVA pathway from plasmid pS-NA
|
This study
|
pS-NA
|
pSTV28 containing mvaE and mvaS from E. faecalis; mvaK1, mvaK2, and mvaD from S. pneumoniae; and idi from E. Coli
|
[19]
|
Strains
|
MG1655
|
E. coli K-12; F−λ−, ilvG−, rfb-50, rph-1
|
ATCC 700926
|
AceCo
|
MG1655 ΔackA-pta, poxB, ldhA, dld, adhE, pps, atoDA
|
Additional file 1: Table S1
|
DH5α
|
E. coli K-12; F−, φ80dlacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17 (rK− mK+), phoA, supE44, λ−, thi-1
|
ATCC 98040
|
MGpPapM
|
MG1655 harboring pT-PaispS and pS-NA
|
This study
|
MGpKpM
|
MG1655 harboring pT-KispS and pS-NA
|
This study
|
MGpPtpM
|
MG1655 harboring pT-PtispS and pS-NA
|
This study
|
MGpsPtpM
|
MG1655 harboring pTS-sPtispS and pS-NA
|
This study
|
MGpsPtM
|
MG1655 harboring pTS-sPtispS-MVA
|
This study
|
ApsPtM
|
AceCo harboring pTS-sPtispS-MVA
|
This study
|