TABLE 1.
Primer names and sequences used for PCR and plasmid construction
| Plasmids | Forward primer (5’→3’) | Reverse primer (5’→3’) |
|---|---|---|
| pTriEGFP | EGFPF (actgccatggTGAGCAAGGGCGAGGA)* | EGFPR (gcatggatccTTGTACAGCTCGTCCAT) |
| MHVnsp15 | mNSP15F (CAACCACAGCGGGTTTTACT) | mNSP15R (GGCTTGCCATAATTCCAGAG) |
| MHVEGFPnsp15 | MHVnsp15F1bamHI (agtcggatccAAGTTTAGAAAATGTA) | MHVnsp15R1 (gactgcggccgcCTGCAAACGAGGATAGAA) |
| F50 | MHVnsp15F50 (agtcggatccAACTAATGTGGCTGTCGAA) | MHVnsp15R1 |
| F100 | MHVnsp15F100 (agtcggatccAGTCTGCAAATACACAGAT) | MHVnsp15R1 |
| F150 | MHVnsp15F150 (agtcggatccACCACAACGTGCCGATTT) | MHVnsp15R1 |
| F200 | MHVnsp15F200 (agtcggatccAAATCCGGGTGGTAATCGCGT) | MHVnsp15R1 |
| F250 | MHVnsp15F250 (agtcggatccAGCAAAATATAGTTTACA) | MHVnsp15R1 |
| F300 | MHVnsp15F300 (agtcggatccATCTAGCATTCATTCGTACTT) | MHVnsp15R1 |
| R50 | MHVnsp15F1bamHI | MHVnsp15R50 (gactgcggccgcGGGGAATGGCGTGTT) |
| R100 | MHVnsp15F1bamHI | MHVnsp15R100 (gactgcggccgcCTTATACGTCGAACTGCA) |
| R150 | MHVnsp15F1bamHI | MHVnsp15R150 (gactgcggccgcGCCTTTAATCATCGACA) |
| R200 | MHVnsp15F1bamHI | MHVnsp15R200 (gactgcggccgcACCTTGTGGGCTCCGGTA) |
| R250 | MHVnsp15F1bamHI | MHVnsp15R250 (gactgcggccgcAATGAACACATCATCAT) |
| R300 | MHVnsp15F1bamHI | MHVnsp15R300 (gactgcggccgcGTCGTATGTCACGAACTCTT) |
| MHVnsp15P101-150 | MHVnsp15F100 | MHVnsp15R150 |
| MHVnsp15P301-374 | MHVnsp15F300 | MHVnsp15R1 |
| mP301-324 | MHVnsp15F300 | MHVnsp15R324 (gactgcggccgcTTAATCAATAACAGTGCACA) |
| mP301-350 | MHVnsp15F300 | MHVnsp15R350 (gactgcggccgcTTAAACATTAACAACCTTA) |
| mP325-350 | MHVnsp15F325 (agtcggatccATTATTGTTAGATGATTT) | MHVnsp15R350 |
| mP350-374 | MHVnsp15F350 (agtcggatccAGATTTTAAAGATTTCCA) | MHVnsp15R1 |
| SARSEGFPnsp15 | SARSnsp15F1 (agtcggatccGAGTTTAGAAAATGTGGCTTA) | SARSnsp15R1 (gactgcggccgcTTGTAGTTTTGGGTAGAA) |
| SARSnsp15P101-156 | SARSnsp15F101 (agtcggatccAGTCTGCACAATGACTGA) | SARSnsp15R156 (gactgcggccgcTTATCCCTTTGAAGGTGTTA) |
| SARSnsp15P273-346 | SARSnsp15F273 (agtcggatccAAGCACAGTGAAAAATTA) | SARSnsp15R1 |
| SARSnsp15minicore | SARSnsp15F287 (agtcggatccATCATCAAAATGTGTGTGT) | SARSnsp15R323 (gactgcggccgcTTAGTCAATTGTAACCTTGA) |
| TGEVEGFPnsp15 | TGEVnsp15F1 (agtcggatccGAGTCTAGAAAATGTGGCTTT) | TGEVnsp15R1 (gactgcggccgcTTGGAGTTGTGGATAGAA) |
| TGEVnsp15P101-148 | TGEVnsp15F101 (agtcggatccAGTGTGTTCCTACACTGAT) | TGEVnsp15R148 (gactgcggccgcTTATAATTTAATGGCACTA) |
| TGEVnsp15P266-339 | TGEVnsp15F266 (agtcggatccAAGTACACTGAAAAGTT) | TGEVnsp15R1 |
| TGEVnsp15minicore | TGEVnsp15F280 (agtcggatccATCTTCTAAGAATGTGT) | TGEVnsp15R316 (gactgcggccgcTTAATCTACAATGACATCCA) |
| SARS/MHV-Domain I | SARSnsp15F1 | SMnsp15R1 (ATCTGTGTATTTGCAGACACCTATTGTAGATACAT) |
| SMnsp15F2 (ATGTATCTACAATAGGTGTCTGCAAATACACAGAT) | SMnsp15R2 (TTTGAAGGTGTTAGGCCTTTAATCATCGA) | |
| SMnsp15F3 (TCGATGATTAAAGGCCTAACACCTTCAAA) | SARSnsp15R1 | |
| TGEV/MHV-Domain I | TGEVnsp15F1 | TMnsp15R1 (ATCTGTGTATTTGCAGACTTGCTTAGTGAAATTT) |
| TMnsp15F2 (AAATTTCACTAAGCAAGTCTGCAAATACACAGAT) | TMnsp15R2 (ATCATTCAAAAGGCCGCCTTTAATCATCGA) | |
| TMnsp15F3 (TCGATGATTAAAGGCGGCCTTTTGAATGAT) | TGEVnsp15R1 | |
| SARS/MHV miniD | SARSnsp15F273 | SMR1 (TGCACACACTCTTACTACTACCTGTTTGCGCATCT) |
| SMF2 (AGATGCGCAAACAGGTAGTAGTAAGAGTGTGTGCA) | SMR2 (AATGAAATTTCAGCATAATCAACATTAACATTAA) | |
| SMF3 (TTAATGTTAATGTTGATTATGCTGAAATTTCATT) | SARSnsp15R1 | |
| TGEV/MHV miniD | TGEVnsp15F266 | TMR1 (ACACACTCTTACTACTTGGATCATCAGCATAT) |
| TMF2 (ATATGCTGATGATCCAAGTAGTAAGAGTGTGT) | TMR2 (ATCTCCATGCCTTACAATCAACATTAACATT) | |
| TMF3 (AATGTTAATGTTGATTGTAAGGCATGGAGAT) | TGEVnsp15R1 | |
| pTriGST | GSTNcoI (atgcccATGgCCCCTATACTAGGTTATT) | GSTNotI (atgcGCGGCCGCcTCGAGTCGACCCGGGAA) |
| pTriGSTnsp15C300 | MHVnsp15F300 | MHVnsp15R1 |
*
The noncoding sequence is indicated in low case letter and the restriction enzyme site is underlined.