TABLE 1.
Primer use | Sequence (5′→3′) | Targeta |
---|---|---|
Oligonucleotides for making targeted galK insertions into pp71 | CGG AAG CGG CCG CGA TCA GCG AGG CCG AAG CCG CCA GCG GCA GCT TTG GTC CTG TTG ACA ATT AAT CAT CGG CA | pp71 C34S GalK FWD |
CGT AGA CGA CCG GCT TCC AGC GAG CCG CCT TCC ACG TTG GTG ATG AGC CGT CAG CAC TGT CCT GCT CCT T | pp71 C34S GalK REV | |
AGG AGA ACA AAT CTC CGC ACG ACA CCG TAG ACC TGA CCG ATC TAA ACA TCC CTG TTG ACA ATT AAT CAT CGG CA | pp71 C94S GalK FWD | |
AGG CGT CGT GGG CCA AAG TTG TTG AGG TCC ACC AGT AGT CGG TCC TGT TCT CAG CAC TGT CCT GCT CCT T | pp71 C94S GalK REV | |
TCG CCT TTC GCG CCG GCG CTA TCC CGC TGA CGT TGG TAG ACG CCC TGG AGC CTG TTG ACA ATT AAT CAT CGG CA | pp71 C218S GalK FWD | |
ATG ATC CAT TGG CCT CGC TCG TCC GTC TCC GTT TTG TGG ATG TAC GTA TTT CAG CAC TGT CCT GCT CCT T | pp71 C218S GalK REV | |
Oligonucleotides for making S-nitrosylation-resistant mutants (galK reversions) | CAG CGA GGC CGA AGC CGC CAG CGG CAG CTT TGG TCG CCT GCA CTC ACA GGT GCT TCG GCT CAT CAC CAA CGT GGA AGG CGG CTC GCT GG | pp71 C34S Mut FWD |
CCA GCG AGC CGC CTT CCA CGT TGG TGA TGA GCC GAA GCA CCT GTG AGT GCA GGC GAC CAA AGC TGC CGC TGG CGG CTT CGG CCT CGC TG | pp71 C34S Mut REV | |
GCA CGA CAC CGT AGA CCT GAC CGA TCT AAA CAT CAA GGG CCG CTC AGT GGT GGG CGA ACA GGA CCG ACT ACT GGT GGA CCT CAA CAA CT | pp71 C94S Mut FWD | |
AGT TGT TGA GGT CCA CCA GTA GTC GGT CCT GTT CGC CCA CCA CTG AGC GGC CCT TGA TGT TTA GAT CGG TCA GGT CTA CGG TGT CGT GC | pp71 C94S Mut REV | |
CGC TAT CCC GCT GAC GTT GGT AGA CGC CCT GGA GCA GCT GGC CTC ATC GGA CCC TAA TAC GTA CAT CCA CAA AAC GGA GAC GGA CGA GC | pp71 C218S Mut FWD | |
GCT CGT CCG TCT CCG TTT TGT GGA TGT ACG TAT TAG GGT CCG ATG AGG CCA GCT GCT CCA GGG CGT CTA CCA ACG TCA GCG GGA TAG CG | pp71 C218S Mut REV | |
Sequencing primers for confirming S-nitrosylation-resistant mutants | CGA CCT CCC ACG AAG ACC C | pp71 C34S Seq FWD |
GTC TAC GGT GTC GTG CGG AG | pp71 C34S Seq REV | |
AGG TGC TTC GGC TCA TCA CC | pp71 C94S Seq FWD | |
CCG CTG GCT CTG AAA GAG GT | pp71 C94S Seq REV | |
CCG CAC GGC TCA CTA CTG G | pp71 C218S Seq FWD | |
GGT GCG GGT ACG GAT TGT GT | pp71 C218S Seq REV | |
Primers for cloning pp71 (WT/C218S) into pCMSeGFP vector | TCG AGA ATT CGC CAC CAT GGA CTA CAA AGA CGA TGA CGA CAA GAT GTC TCA GGC ATC GTC CTC GC | EcoRI-Kozak-Flag-pp71 FWD |
AGC TGA ATT CCT AGA TGC GGG GTC GAC TGC GTG GGG TGC TGG AAG TGG AAG CGG TG | EcoRI-Kozak-Flag-pp71 REV | |
Primers for cloning pp71 (WT/C218S) into pLXSN vector | TCG AGA ATT CGC CAC CAT GTC TCA GGC ATC GTC CTC GCC CGG TGA GGG ACC CTC GTC GGA | EcoRI-Kozak-pp71 FWD |
AGC TGA ATT CCT AGA TGC GGG GTC GAC TGC GTG GGG TGC TGG AAG TGG AAG CGG TG | EcoRI-Kozak-pp71 REV | |
Real-time PCR primers | CAG CAA TTT TCA GTG TCA GAA GC | IFN-β1 FWD |
TCA TCC TGT CCT TGA GGC AGT | IFN-β1 REV | |
GAA TGG GTT TGC TAG AAT GTG ATA | IL-1β FWD | |
CAG ACT AGG GTT GCC AGA TTT AAC | IL-1β REV | |
ATG GTT TTG CAG GCT TTG ATG | UL122 FWD | |
ACC TGC CCT TCA CGA TTC C | UL122 REV | |
CCC TCG GCT TCT CAC AAC AAT | UL54 FWD | |
CGA GTT AGT CTT GGC CAT GCA T | UL54 REV | |
GGT TTC TGG CTC GTG GAT GTC G | UL32 FWD | |
CAC ACA ACA CCG TCG TCC GAT TAC | UL32 REV | |
ACC CAC TCC TCC ACC TTT GAC | GAPDH FWD | |
CTG TTG CTG TAG CCA AAT TCG T | GAPDH REV | |
ACC ACT ACC TGA GCA CCC AGT C | eGFP FWD | |
GTCCATGCCGAGAGTGATCC | eGFP REV | |
CCC CTT CCA TCA CAT TGC A | mdm2 FWD | |
AGT TTG GCT TTC TCA GAG ATT TCC | mdm2 REV |
FWD, forward primer; REV, reverse primer.