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. 2020 Aug 17;94(17):e00033-20. doi: 10.1128/JVI.00033-20

TABLE 1.

Oligonucleotides used in this study

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
a

FWD, forward primer; REV, reverse primer.