Table 1.
Primers used for recombinant A33 construction by site-directed mutagenesis of the A33RVACVgene
| Mutation | Primer sequencea |
|---|---|
| D115A |
ACATTCAGCATACCAGTTATTCTCGGATGCTAAAGCAAATTGCACTGCGG |
| Y116A |
ACATTCAGACGCACAGTTATTCTCGGATGCTAAAGCAAATTGCACTGCGG |
| Q117A |
ACATTCAGACTACGCATTATTCTCGGATGCTAAAGCAAATTGCACTGCGG |
| L118A |
ACATTCAGACTACCAGGCATTCTCGGATGCTAAAGCAAATTGCACTGCGG |
| S120A |
ACATTCAGATTACCAGTTATTCGCTGATGCTAAAGCAAATTGCACTGCGG |
| N125A |
ACATTCAGACTACCAGTTATTCTCGGATGCTAAAGCAGCATGCACTGCGG |
| E129A |
ACATTCAGACTACCAGTTATTCTCGGATGCTAAAGCAAATTGCACTGCGGCATCATCAAC |
| D115A,Q117A |
ACATTCAGCTTACGCATTATTCTCGGATGCTAAAGCAAATTGCACTGCGG |
| D115A,N125A |
ACATTCAGCTTACCAGTTATTCTCGGATGCTAAAGCAGCATGCACTGCGG |
| Q117A,N125A |
ACATTCAGACTACGCATTATTCTCGGATGCTAAAGCAGCATGCACTGCGGAATCATCAAC |
| L118A,E129A | ACATTCAGACTACCAGGCATTCTCGGATGCTAAAGCAAATTGCACTGCGGCATCATCAAC |
aAll primers listed in 5′-3′ direction with the position of the mutation(s) underlined. A common reverse primer was used for all reactions:
AATATATAACAAGAACCCTGGTAATAGAGACCATTACAGCTTTC.