TABLE 5.
List of primers used to generate the levansucrase inactivation construct and for qRT-PCR
| Primer name | Primer sequence (5′→3′)a |
|---|---|
| Levansucrase inactivation construct | |
| LevFragA_fwd | TGGGGATCCTTGAAGTTTAACAAATGGTTCAGTAAAGC |
| LevFragA_rev | CCTCCTAAACAGTTAGGACGGAACCTCATATTTCTCTTTGCC |
| LevFragB_fwd | ACTCTTATTTTTTTAATATTGTTTCATAGTGGCAATAACGTAGTCG |
| LevFragB_rev | GGGCTCGAGTTATTTCTTTCCATACTCATTTGGAG |
| Erm_fwd | TAACTGTTTAGGAGGACTGATAATATGAACAAAAATATAAAATATTCTCAAAAC |
| Erm_rev1 | TAAAAAAATAAGAGTTACCATTTATTACCTCCCGTTAAATAATAGATAAC |
| Erm_rev2 | GCCACTATGAAACAATATTAAAAAAATAAGAGTTACCATTTATTATTTCC |
| qRT-PCR | |
| Lev_Fwd | GTACAGCAAAGCGTCGGAAT |
| Lev_Rev | CCGGTTTCTGTTCCTGTGTT |
| Erythromycin_Fwd | GGTTGCTCTTGCACACTCAA |
| Erythromycin_Rev | CTGTGGTATGGCGGGTAAGT |
| Polysaccharide polymerase_Fwd | GCGTTCGTCGGTTTATCACT |
| Polysaccharide polymerase_Rev | GAATGCAGCCCTAGAACCTG |
| Flippase_Fwd | CGTTCCAAGCAGAAAGGAAG |
| Flippase_Rev | AGACAACAGCGAACCTGCTT |
| Exopolysaccharide biosynthesis protein_Fwd | GGTCACATTCTGGCCTGTCT |
| Exopolysaccharide biosynthesis protein_Rev | CTAAACAGCTTCGCCTTTGG |
| 16S rRNA_Fwd | GGCTTTCCAGCTACCTGTTG |
| 16S rRNA_Rev | ACGGCGTCTTCAAAGGAGTA |
a
Underlined sequences represent either restriction sites or RBS.