Table 1 Primers used for ΔMycP3ms generation, MycP3 complementation and RT-qPCR assay
Experiment | Gene related | Primer name | Primer sequence | Restriction site (underlined) |
|---|---|---|---|---|
ΔMycP3ms Generation | mycP 3 | UP forward | 5′-AAGCTTTCCCACGCACATCG-3′ | HindIII |
UP reverse | 5′-CTCG AGATCACCTGTCGAGCACG-3′ | XhoI | ||
DOWN forward | 5′-CTCGAGATGACCGCCCGGATAGC-3 | XhoI | ||
DOWN reverse | 5′-GGATCCCCGGTCTCGGTGAC-3′ | BamHI | ||
ΔMycP3ms Construct verification | mycP 3 | mycP 3 screening forward | 5′-GCTCAACCCGAAGATC GCCTC-3′ | N/A |
mycP 3 screening reverse | 5′-AGGAACATGCCTTTCCACCAGG-3′ | N/A | ||
MycP3 Complementa-tion | mycP 3 | pr1 forward | 5′-CCATGGGACGCTGAACGAGTGTTTAC-3′ | NcoI |
pr1 reverse | 5′-GACGCCCAGACTCTTGTGGATCACATCGCGGTCGACCCGGGGCG-3′ | N/A | ||
mycP 3 -pr1 forward | 5′-CGCCCCGGGTCGACCGCGATGTGATCCACAAGAGTCTGGGCGTC-3′ | N/A | ||
mycP 3 reverse | 5′-AAGCTTTCATGTGGTCTTGTCCTTCC-3′ | HindIII | ||
pr2 forward | 5′-CCATGGACGTGGGACGGCGACGA GAATC-3′ | NcoI | ||
pr2 reverse | 5′-GACGCCCAGACTCTTGTGGATCACGACTGTTTCC TTTCGAAGGTGGTG-3′ | N/A | ||
mycP 3 -pr2 forward | 5′-CACCACCTTCGAAAGGAAAC AGTCGTGATCCACAAGAGTCTGGGCGTC-3′ | N/A | ||
RT-qPCR assay | sigA | sigA forward | 5′-GGGCGTGATGTCCATCTGCT-3′ | N/A |
sigA reverse | 5′-GTATCCCGGTGCATGGTC-3′ | |||
mycP 3 | mycP 3 forward | 5′-GGATCATCGCGTTCGTGGGTAC-3′ | ||
mycP 3 reverse | 5′-GTCTTGTCCTTCCGACGGTAGG-3′ | |||
eccE 3 | eccE 3 forward | 5′-GAGCCGTTGTTGACGGTTTG-3′ | ||
eccE 3 reverse | 5′-GTTCGGTCGACAACGGGTTC-3′ |