Table 1.
Oligonucleotides and plasmids generated and used in the study.
| Name | Sequence/Description |
|---|---|
| OLIGONUCLEOTIDES | |
| pMV3′ | GCCTGGCAGTCGATCGTACG |
| pUX1-NheI | ACGGCATGGACGAGCTGTAC |
| pMV(NheI-AflII)a | CCGCGGTGATCAGCTTAAGCCAACAAAGCGAC |
| mmpL4a_Fw_SpeI | TGTGACTAGTCAGATGGGGAAGGTCTTTCA |
| mmpL4a_Rev_BamHI_100 | CAAGAAGGATCCCGTAATACTTGTGCGCATCGTCTCC |
| mmpL4a_Rev_BamHI_500 | GAGAGGATCCCGGTGAACAACAGGATGATG |
| mmpL4a_Rev_BamHI_1000 | GAGAGGATCCCGGTGTAGCTGGGGTTGTAT |
| mmpL4a_Rev_BamHI_1500 | GTGTGGATCCTGTTTGAGCATGTCGTCCAT |
| mmpL4a_conf_left | CTTCCGTGGTCCGTCAAAT |
| mmpL4a_conf_right | CATTCGTGAGACCAGCAACA |
| mmpS4_Fw_BamHI | GAGTAGGATCCATGCGTCTGTGGATTCCGCTG |
| mmpS4_Rev_AflII | GAGTACTTAAGCATGATGCTTCCCACCGCG |
| mmpS4_conf_left | GGATACCCAGTGGCTTGAAA |
| mmpL4a-mmpL4b_left | TGCGTCTGTGGATTCCGCTG |
| mmpL4a-mmpL4b_right_HindIII | GAGAGAAAGCTTAGTACGTCATCCCGGTGTTC |
| PLASMIDS | |
| pMV261 | Multicopy E. coli- mycobacterium shuttle vector, kanr (Stover et al., 1991) |
| pMV261-AflII | Variant of pMV261 in which the NheI site of the polylinker was changed to an AflII by site-directed mutagenesis, kanr |
| pMV306-zeo-mmpL4a | Integrative E. coli- mycobacterium shuttle vector, zeor, contains M. abscessus subsp. abscessus mmpL4a under the control of the hsp60 promoter (Bernut et al., 2016) |
| pUX1 | Plasmid produced by ligating the colE1 origin and kanr cassettes-containing XmnI fragment of pMV261-AflII to the blunted SpeI fragment from pTEC27 (Takaki et al., 2013) containing the tdTomato and hygr cassettes. |
| pUX2 | Plasmid produced by ligating the colE1 origin and kanr cassettes-containing XmnI fragment of pMV261-AflII to the blunted SpeI fragment from pTEC15 (Takaki et al., 2013) containing the mWasabi and hygr cassettes. |
| pUX3 | Plasmid produced by replacing the hygr cassette flanked by NheI and BamHI sites of pUX1 with the acetamidase inducer elements of pSD26 (Daugelat et al., 2003). |
a
Underlined are restriction enzyme recognition sequences.