Strains | Â | Source |
T. aotearoense SCUT27 | Wild type strain | [11] |
DH5α | E. coli cloning strain, F-endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZ∆M15 ∆(lacZYA-argF)U169, hsdR17(r K -m K +), λ– | Invitrogen |
LA1002 | As SCUT27, but ∆pta, ∆ack | This study |
Plasmids | Description | Source |
pBluescript II SK(+) | Standard cloning vector, f1 ori; AmpR; | Stratagene |
pBlue-aph | Derived from pBluescript II SK(+), with kanamycin expression cassette | This study |
pBlue-pta-aph | Derived from pBlue-aph, with partial phosphotransacetylase (pta) gene upward of kanamycin gene | This study |
pPuKAd | Homologous recombination plasmid derived from pBlue-pta-aph, with partial acetate kinase (ack) gene downward of kanamycin gene | This study |
Primers | Sequence 5′ → 3′) a | Application |
pta-F | AACTAGGTACCAGCGCTGTACGAAATTGCCACTC | Forward primer for pta |
pta-R | GTACTGAATTCCACCCATTCCTTGTGTTATAGG | Reverse primer for pta |
ack-F | GAGCGGATCCGCATAGAATTAGCTCCACTGC | Forward primer for ack |
ack-R | TGACTGCGGCCGCCGACGCCTCCCATAGCTG | Reverse primer for ack |
Prob-F | TATTAAGACCTGCATTTCAGAT | Forward primer for hybridization probe |
Prob-R | CATTTGCCTTAGCTAACCTC | Reverse primer for hybridization probe |