Table 1.
Plasmids and primers
| Plasmids/primers | Properties/sequence | Use | Origin |
|---|---|---|---|
| pCMV-Myc-TLR2 | Eukaryotic expression plasmid encoding TLR2 | Transfection | [43] |
| AU1-MyD88 full | Eukaryotic expression plasmid encoding the full-length human MyD88 | Transfection | [44] |
| DN-AU1-MyD88 | Eukaryotic expression plasmid encoding TIR domain (152–296 aa) of human MyD88; acts as dominant negative on MyD88 signaling | Transfection | [44] |
| NF-κB-luc | Reporter of the activity of transcription factor NF-κB | Transfection | Stratagene |
| Myc-DDK-TIRAP | Eukaryotic expression plasmid encoding the adaptor protein TIRAP | Transfection | OriGene |
| pEGFP-TirS | Eukaryotic expression plasmid encoding TirS | Transfection | This work |
| pEGFP-C2 | Eukaryotic expression vector | Transfection | BD Biosciences |
| pCMV | Eukaryotic expression vector | Transfection | Clontech |
| pcDNA3 | Eukaryotic expression vector | Transfection | Invitrogen |
| PKOR1 | S. aureus shuttle vector | Making isogenic mutant | [45] |
| pCM29 | GFP fluorescence expression vector | Reporter | [46] |
| tirS prev For | 5′-CAGTCTTACCTGCTCGATTC-3′ | RT-PCR | This work |
| tirS prev Rev | 5′-CTTACGCACATCAATAACGA-3′ | RT-PCR | This work |
| gyrA prev For | 5′-GTCAAAATCTGCAAAAATAGCTAG-3′ | RT-PCR | This work |
| gyrA prev Rev | 5′-GTCAAAATCTGCAAAAATAGCTAG-3′ | RT-PCR | This work |
| tirS EcoRI For | 5′-AATCTAGAATTCGAGGTATTATATGTCAG-3′ | Cloning | This work |
| tirS BamHI Rev | 5′-ATTGTTCTCGGATCCTTCCCTCTTTGCTTTTAAAG-3′ | Cloning | This work |
| Up For tirS + attB1 | 5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTCGCTAGGTTG GGCTTTTCCACAT-3′ | Generating fusion construct | This work |
| Up Rev tirS | 5′-AAAGTAGATAACCAATACTATTAATAATACCTCGCTTTTT ATAATC-3′ | Generating and sequencing fusion construct | This work |
| Down For tirS | 5′-GATTATAAAAAGCGAGGTATTATTAATAGTATTGGTTATC TACTTT-3′ | Generating and sequencing fusion construct | This work |
| Down Rev tirS + attB2 | 5′-GGGGACCACTTTGTACAAGAAAGCTGGGTCAGTTACTCC CGCTTCTGTTAATG-3′ | Generating fusion construct | This work |
| pKOR1 For | 5′-AGCTCCAGATCCATATCCTTC-3′ | Sequencing fusion construct | This work |
| pKOR1 Rev | 5′-CACACAGGAAACAGCTATGAC-3′ | Sequencing fusion construct | This work |
| tirS KO confirm For | 5′-GCTTCGAGAGTGGTTAGAC-3′ | Isogenic mutant confirm | This work |
| tirS-CL-For | 5′-GCGGAATTCATGTCAGTATTAGAAACTAAATTAAAAAGTC-3′ | Complementation | This work |
| tirS-CL-Rev | 5′-GCGAAGCTTCTAATTCTTAGAATTAACGATTACTTG-3′ | Complementation | This work |
| pDC123 | 5′-GCGGAATTCATGTCAGTATTAGAAACTAAATTAAAAAGTC-3′ | Shuttle vector used for complementation | [45] |
| tirS KO confirm Rev | 5′-GGTTATCATCAAATGAGCTACCTG-3′ | Isogenic mutant confirm | This work |
| tirS Int For | 5′-TTAATCTTCAAAAAGAGCAGTCTAGG-3′ | Isogenic mutant confirm | This work |
| tirS Int Rev | 5′-GGGTTGTATGCACGTACATCTTCAAC-3′ | Isogenic mutant confirm | This work |
| tirS-prom For | 5′-CGCCTGCAGGCTCCACAGTTTGTTCATCCTGAT-3′ | Generating reporter construct | This work |
| tirS-prom Rev | 5′-CGCGGTACCCGTTTGTTCCCTTTTATATTGTATATCTTATC-3′ | Generating reporter construct | This work |