Skip to main content

Table 1 Strains and plasmids used in this study

From: Biological conversion of methane to putrescine using genome-scale model-guided metabolic engineering of a methanotrophic bacterium Methylomicrobium alcaliphilum 20Z

Strain or plasmid Characteristics Reference or source
Strains
 E. coli DH5α F– endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG purB20 ϕ80dlacZΔM15 Δ(lacZYAargF)
U169, hsdR17(rK–mK +), λ–
Solgent
 M. alcaliphilum 20Z Wild-type strain 20Z DMSZ
 20ZE Evolved putrescine-tolerant strain M. alcaliphilum 20Z This study
  20ZES1 Evolved putrescine-tolerant strain ΔspeE1 This study
  20ZE3 Evolved putrescine-tolerant strain ΔldhΔackΔspeE1 This study
  20ZE4A Evolved putrescine-tolerant strain ΔldhΔackΔspeE1::argDJ This study
Plasmid
 pAWP89 pAWP78 containing dTomato driven by pTac promoter [49]
 pCM433KanT A broad host-range sacB-based vector for unmarked allelic exchange, Kmr [13]
 pCSE1 pCM433kanT containing the deletion construct of speE1 This study
 pCM433ldh pCM433kanT containing the deletion construct of ldh [16]
 pCM433ack pCM433kanT containing the deletion construct of ack [16]
 pCM351 pCM184 with aaaC1 from pCM350; allelic exchange vector [50]
 pCAR2 pCM351 containing two flanking regions to replace the speE2 by pTac-argDJ fragment This study
 pACE pAWP89 carrying speC (E. coli) and potE driven by tac promoter This study
 pACO pAWP89 carrying speC (M. trichosporium OB3b) driven by tac promoter This study