Strain, plasmid and primers | Genotype or phenotype |
---|---|
Serratia sp. | |
 LCN16 WT | WT resistant to ampicillin and erythromycin. |
 LCN16 oxyR::int(614) | Knockout mutant of oxyR with group II intron inserted in the position 614; Resistant to gentamicin and kanamycin. |
 LCN16 katA::int(808) | Knockout mutant of katA with group II intron inserted in the position 808; Resistant to gentamicin and kanamycin. |
Escherichia coli | |
 DH5α | Competent cells (Sigma-Albrich) |
Plasmids | |
 pAR1219 | TargeTron vector with chloramphenicol resistant |
 pAR1219-ΩGm | Constructed vector with gentamicin resistant gene aacC |
 pACD4K-C | TargeTron vector; kanamicin RAM marker (for chromosomal insertion) and chloramphenicol resistance (plasmid propagation) |
 pACD4K-C_oxyR | TargeTron vector with intron RNA retarget for oxyR gene |
 pACD4K-C_kat | TargeTron vector with intron RNA retarget for kat gene |
 pBK-miniTn7-ΩGm | Tn7 plasmid constructed with gentamicin resistant gene aacC |
PCR Primers (5′-3′) | |
 aacC1_IfFor | CATACTCTTCCTTTTTCAATATTATTG |
 aacC1_IfRev | TAACTGTCAGACCAAGTTTACTC |
 oxyR_614|615 s-IBS | AAAAAAGCTTATAATTATCCTTAGGAAGCTGGTCAGTGCGCCCAGATAGGGTG |
 oxyR_614|615 s-EBS1d | CAGATTGTACAAATGTGGTGATAACAGATAAGTCTGGTCACTTAACTTACCTTTCTTTGT |
 oxyR_614|615 s-EBS2 | TGAACGCAAGTTTCTAATTTCGATTCTTCCTCGATAGAGGAAAGTGTCT |
 oxyR_KO_chkFor | CGTGGTCTGGAGGGAAACAA |
 oxyR_KO_chkRev | CATAACGACTGCGCAATGGG |
 katA_808|809a -IBS | AAAAAAGCTTATAATTATCCTTATAATCCGGATTTGTGCGCCCAGATAGGGTG |
 katA_808|809a EBS1d | CAGATTGTACAAATGTGGTGATAACAGATAAGTCGGATTTGCTAACTTACCTTTCTTTGT |
 katA_808|809a -EBS2 | TGAACGCAAGTTTCTAATTTCGGTTGATTATCGATAGAGGAAAGTGTCT |
 katA_KO_chkFor | GGTGAAGTTCCATTTCCGCTGC |
 katA_KO_chkRev | GGGTTCACCGCTACCTGTTCAAC |
RT-qPCR primers (5′-3′) | |
 gyrA_For | TTATCTCCCTGATTGTGCCA |
 gyrA_Rev | CATTACGCTCGCTCACCTTA |
 oxyR_For | TTTAGAGTACCTGGTCGCCTTG |
 oxyF_Rev | ATCACACCCAGTTCGTCTTCC |
 katA_For | CCAGATTATGCCTGAACACG |
 katA_Rev | TGCAGTTCGAAGAAACCAAC |
 katG_For | AGCGGTAAGCCAAATACACC |
 katG_Rev | AATCGAAGTCAGGGTCCATC |