Oligonucleotides used in confirming successful transduction of serine
hydrolase mutants into S. aureus Newman can be found inTable S5 . In brief,
transposon mutagenesis was used to generate insertion mutations in each
NWMN_0169, NWMN_0262, NWMN_0748, NWMN_1210, NWMN_1683, NWMN_2092, NWMN_2350,
NWMN_2379, NWMN_2434, NWMN_2480, NWMN_2528, and NWMN_2569. Homologs of the
aforementioned genes were first identified through the Basic Local Alignment
Search Tool (BLASTn) in S. aureus USA300 FPR3757 (GeneBank ID
CP000255.1), and corresponding transposon mutants were identified and confirmed
in the Nebraska Transposon Mutant Library30 (link) through PCR. The mutations were mobilized into
S. aureus Newman using transduction with phage ϕ85.
Successful transductants were confirmed through PCR and assessed for hemolysis
relative to wild-type S. aureus Newman and USA300 by streaking
on BD Trypticase Soy Agar II with 5% Sheep’s Blood (BD).
hydrolase mutants into S. aureus Newman can be found in
transposon mutagenesis was used to generate insertion mutations in each
NWMN_0169, NWMN_0262, NWMN_0748, NWMN_1210, NWMN_1683, NWMN_2092, NWMN_2350,
NWMN_2379, NWMN_2434, NWMN_2480, NWMN_2528, and NWMN_2569. Homologs of the
aforementioned genes were first identified through the Basic Local Alignment
Search Tool (BLASTn) in S. aureus USA300 FPR3757 (GeneBank ID
CP000255.1), and corresponding transposon mutants were identified and confirmed
in the Nebraska Transposon Mutant Library30 (link) through PCR. The mutations were mobilized into
S. aureus Newman using transduction with phage ϕ85.
Successful transductants were confirmed through PCR and assessed for hemolysis
relative to wild-type S. aureus Newman and USA300 by streaking
on BD Trypticase Soy Agar II with 5% Sheep’s Blood (BD).
