Genetic Confirmation of the Role of Sulfopyruvate Decarboxylase in Coenzyme M Biosynthesis in Methanococcus maripaludis

Table 1

List of strains, plasmids, and primers.

Strains	Description	Source
M. maripaludis S2	Wild type	[4]
M. maripaludis S201	comE::Tn5<KAN-2-pac>	This work
M. maripaludis S202; M. maripaludis S203	comE::Tn5<KAN-2-pac>; Strain S201 complemented with pMEVII-comE	This work; This work
E. coli EC100	F mcrA Δ(mrr-hsdRMS  mcrBC) Φ80dlacZΔM15 ΔlacX74  recA1  endA1  araD139  Δ(ara, leu)7697  galU  galK  𝜆  rpsI (str)  nupG	Epicentre
Plasmids
pUC18	E. coli plasmid cloning vector	GenScript
pComE	Plasmid pUC18 containing the genetic region of gene comE	This work
PComET1 and 2	pComE containing Tn5 transposon insertions in comE gene	This work
pMEVI and pMEVII	Shuttle plasmids for M. maripaludis	[5]
pMEVII-comE	Plasmid pMEVII containing gene comE in trans	This work
Primers
RegcomEF	5′-AAAAAAGGATCCCGGATCTGACCCATACAATAGAG	This work
RegcomER	5′-AAAAAATCTAGAATGGATGGATTG GCAGTTTC	This work
ME-Plus9-3′	5′-CTGTCTCTTATACACATCTCAACCATCA	Epicentre
ME-Plus9-5′	5′-CTGTCTCTTATACACATCTCAACCCTGA	Epicentre
KAN-2RP-1out2	5′-GCAATGTAACATCAGAGATTTTGAC	[6]
MMP1688R	5′-CGAATGGATTCTTTTGAACTTTT	This work
comEF	5′-TTGCGTTCATAAATCTGTGTTT	This work
comER; comEexpF; comEexpR	5′-AATGGAATACGTGACCGATG; 5′-ATATATATGCATGGAACTTACGAGATA; 5′-ATATATAGATCTTTTATTTTTTTATTGCG	This work; This work; This work
M13R	5′-CAGGAAACAGCTATGACC