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Table 1 Strains, plasmids and primers used in this study

From: Retinoid production using metabolically engineered Escherichia coli with a two-phase culture system

Strains, plasmids and primers

Description

Reference or source

E. coli strains

  

   MG1655

K12, Wild type

 

   DH5α

F-, ϕ 80dlac ZΔ M15, Δ(lacZYA-argF)U169, deoR, recA 1 endA 1, hsdR 17(rK- mK+), phoA, supE 44, λ-, thi-1, g yrA96, rel A1

 

   XL1-Blue

hsdR 17, supE 44, recA 1, endA 1, gyrA 46, thi, relA 1, lac/F'[proAB+, lacIq, lacZΔ M15::Tn10(tetr)]

 

   S17-1

recA pro hsdR RP4-2-Tc::Mu-Km::Tn7

 

   BL21(DE3)

F-, ompT, hsdSB(r B -m B -), gal (λc I 857, ind 1, Sam 7, nin 5, lac UV5-T7gene 1), dcm (DE3)

 

Plasmids

  

   pBluescript

Plac cloning vector, ColE1 origin, lacZ, Ampr

Stratagene

   pSTV28

Plac expression vector, pACYC184 origin, lacZ, Cmr

Takara

   pTrc99A

Ptrc expression vector, pBR322 origin, lacIq, Cmr

Amersham Bioscience

   pT-HB

pTrc99A containing crtE, crtB, and crtI from P. agglomerans, crtY from P. ananatis, and ipiHP1 from H. pluvialis

[15]

   pT-DHB

pT-HB containing dxs from E. coli

[15]

   pT-HBblh

pT-HB containing blh from Halobacterium sp. NRC-1

This study

   pT-HBbrp

pT-HB containing brp from Halobacterium sp. NRC-1

This study

   pT-HBbrp2

pT-HB containing brp2 from N. pharaonis

This study

   pT-HBBcmo1

pT-HB containing Bcmo1 from M. musculus

This study

   pT-HBSR

pT-HB containing the codon-optimized blh gene (SR) from uncultured marine bacterium 66A03

This study

   pT-DHBSR

pT-DHB containing the codon-optimized blh gene (SR) from uncultured marine bacterium 66A03

This study

   pS-NA

pSTV28 containing mvaE and mvaS from E. faecalis; mvaK1, mvaK2, and mvaD from S. pneumoniae; and idi from E. coli

[13]

Primersa

  

   blhE-F

5'-GGAATTC AGGAGGTGTTCGGC ATG CCACACGG-3'

 

   blh-R

5'-GACTAG TTA GAGGACGCCCTGCACGCGGTC-3'

 

   brpE-F

5'-GGAATTC AGGAGGTATTCAT ATG AGCAATAGGTC-3'

 

   brp-R

5'-GACTAG TTA TGGGACGTACCAGATGCCG-3'

 

   brp2E-F

5'-GGAATTC AGGAGGCCGAGT ATG AGTAACGCGTC-3'

 

   brp2-R

5'-GACTAGTT A TGCTCCGGGTCGCCAGAG-3'

 

   Bcmo1E-F

5'-GGAATTC AGGAGCGGTTCC ATG GAGATAATATTTG-3'

 

   Bcmo1-R

5'-GACTAG TTA AAGACTTGAGCCACCATG-3'

 

   SR-F

5'-GACTAGTGAATTC AGGAGGTAATAAAT ATG G-3'

 

   SR-R

5'-CACTAG TTA GTTTTTGATTTTG-3'

 
  1. a Template binding regions are indicated with bold letters, start and stop codons are underlined, and restriction sites are double underlined.
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Microbial Cell Factories

ISSN: 1475-2859

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