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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Virology. 2014 Jul 16;0:351–362. doi: 10.1016/j.virol.2014.06.001

Table 2.

Primers used to make the minor structural protein knockout mutants

ORF(gene) Primer sequence (5′ to 3′)
ORF2a′ (GP2′) F CCCTCTCCTCTGcTGAGTTTCTGTCCAGG
R CCTGGACAGAAACTCAgCAGAGGAGAGGG
ORF2b′ (E′) F CACTTCTCCTCCTCCTTCAAGTACTAcGATCACACCCACATTCATACTG
R CAGTATGAATGTGGGTGTGATCgTAGTACTTGAAGGAGGAGGAGAAGTG
ORF3′ (GP3′) F GAAGGAGAGTTCTGTACCcTGGTTTCAACTTGGTTTC
R GAAACCAGATTGAAACCAgGGTACAGAACTCTCCTTC
ORF4′ (GP4′) F CCTAACAATCAAGCAATCcTGGAACGAGACAATGATG
R CATCATTGTCTCGTTCCAgGATTGCTTGATTGTTAGG
ORF2a (E) F GCCTTCCACCAcGCTGTCCACGAGCTCC
R GGAGCTCGTGGACAGCgTGGTGGAAGGC
ORF2b (GP2) F CGATCTTTAGAGCATTCTATTCTTAgTGGGTTCTATACTCACCCACATC
R GATGTGGGTGAGTATAGAACCCAcTAAGAATAGAATGCTCTAAAGATCG
ORF3 (GP3) F CAGTTATCTCACTAACCCcTGGATGTCCGTGG
R CCACGGACATCCAgGGGTTAGTGAGATAACTG
ORF4 (GP4) R CCAGTTTGATAACGAGACATTcTGCTACATTGCAAACCTTAC
F GTAAGGTTTGCAATGTAGCAgAATGTCTCGTTATCAAACTGG
Effects of the mutations on an overlapping ORF
CTCTGcTGAGT (ORF2a′): ATG to CTG (knock out start codon)
TACTAcGATCA (ORF2b′): ATG to ACG (knock out start codon)
TACTAcGATCA (ORF2a′): TAT to TAC (Tyrosine to Tyrosine)
GTACCcTGGTT (ORF3′): ATG to CTG (knock out start codon)
GTACCcTGGTT (ORF2a′): CCA to CCC (Proline to Proline)
CAATCcTGGAA (ORF4′): ATG to CTG (knock out start codon)
CAATCcTGGAA (ORF3′): TCA to TCC (Serine to Serine)
CACCAcGCTGT (ORF2a): ATG to ACG (knock out start codon)
CACCAcGCTGT (ORF2b): CAT to CAC (Histidine to Histidine)
TCTTAgTGGGT (ORF2b): ATG to GTG (knock out start codon)
TCTTAgTGGGT (ORF4′): TAA to TAG (STOP codon to STOP codon)
AACCCcTGGAT (ORF3): ATG to CTG (knock out start codon)
AACCCcTGGAT (ORF2): CCA to CCC (Proline to Proline)
ACATTcTGCTA (ORF4): ATG to CTG (knock out start codon)
ACATTcTGCTA (ORF3): TTA to TTC (Leucine to Leucine)
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