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Table 3 Strains and plasmid used in this study

From: Co-production of acetoin and succinic acid by metabolically engineered Enterobacter cloacae

Name

Relevant genotype

Source and ref

Strains

 DH5α

Fφ80 lac ZΔM15 Δ(lacZYA-arg F) U169 endA1 recA1 hsdR17(rk,mk+) supE44λ thi -1 gyrA96 relA1 phoA, Recipient in transformations

Sangon Biotech

 S17-1 λpir

RP4-2(Km::Tn7,Tc::Mu-1) pro-82 LAMpir recA1 endA1 thiE1 hsdR17 creC510; conjugative strain able to host -pir-dependent plasmids

Zoman Biotechnology

 E. cloacae CICC 10011

Wild type

CICC

 EC∆budC

E. cloacae CICC 10011 budC disruption mutant strain

This study

 EC∆ldhA

E. cloacae CICC 10011 ldhA disruption mutant strain

This study

 EC∆budC∆ldhA

E. cloacae CICC 10011 budC and ldhA disruption mutant strain

This study

Plasmid

 pGEM-T Easy Vector

Cloning vector, Apr

Promega

 pToriR6K

pGEM-T Easy Vector with 0.6-kb oriR6K fragment, Apr

This study

 pTBCup

pGEM-T Easy Vector with 0.5-kb budC upstream fragment, Apr

This study

 pTBCdown

pGEM-T Easy Vector with 0.5-kb budC downstream fragment, Apr

This study

 pTLAup

pGEM-T Easy Vector with 0.5-kb ldhA upstream fragment, Apr

This study

 pTLAdown

pGEM-T Easy Vector with 0.5-kb ldhA downstream fragment, Apr

This study

 pK18mobSacB

oriT(RP4) sacB lacZα Plac Pmbi; mobilization and counterselection, Kanr

BCRC

 pRL27

Mini-Tn5 transposon (oriR6K) delivery vector, Kanr

[33]

 pKR6K

R6K replicon; gene replacement vector, Kanr

This study

 pKΔbudC

pKR6K derivative, carries a 771 bp deletion of budC, Kanr

This study

 pK∆ldhA

pKR6K derivative, carries a 990 bp deletion of ldhA, Kanr

This study

  1. CICC, The China Center of Industrial Culture Collection, China; BCRC, The Bioresource Collection and Research Centre, FIRDI, Taiwan