TABLE 1.
Primers used for PCR amplification and their applications in this investigation
| Primer | Sequence | Application | Template | Annealing temp (°C) |
|---|---|---|---|---|
| P042 | 5′-GAAAATCTCATCCTCCGGGGC | Screening recombinant clones and mtuA sequencing | pCALnFLAG-rmtuA plasmids | 55 |
| P043 | 5′-GGATCTAAGCTTGAGCTCGAG | |||
| P064 | 5′-GGATCTAAGCTTGAGCTCGAG | ISS1 primers for Vectorette PCR amplification | Genomic S. uberis 0140J Vectorette libraries | 52 |
| P082 | 5′-CCAACAGCGACAATAATCACATC | |||
| P182 | 5′-CTTGGAGCTTGTTCAGTAGG | PCR amplification of mtuA open reading frame | Genomic S. uberis 0140J | 50 |
| P183 | 5′-TCCATTTCATCATTGCATAA | |||
| P204 | 5′-GAGGAGGAGAAGTGTTCAGTAGGAAATGGTAGA | Ligation-independent cloning expression of mtuA | Genomic S. uberis 0140J | 50 |
| P185 | 5′-GGAACAAGACCCGTATGTGATATTGTGCTTTATTTT |