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. 2009 Dec 18;76(4):1044–1052. doi: 10.1128/AEM.02497-09

TABLE 1.

T. kodakarensis strains and plasmids used and/or constructed in this study

Strain or plasmid Origin and/or genetic featuresa Source or referencef
T. kodakarensis strains
    KW128 ΔpyrF; ΔtrpE::pyrF 21
    TS372 KW128 with trpE-PhmtB-TK1761 insertion 18
    TS416 TS372 with nonsense mutation at codon 3 of TK1761 18
    TS502 KW128 with trpE-PhmtB-(His)10TK1827 insertion This study
    TS503 KW128 with trpE-PhmtB-TK1827 insertion with TAG at codon 5 This study
    TS517 KW128 with ΔTK0664b This study
    TS521 KW128 with ΔTK0664::trpE This study
    TS538 TS517 with ΔTK1761-TK1762-TK1763 This study; Fig. S2
    TS541 TS517 with ΔTK1827 This study; Fig. S2
    TS1079 TS541 with ΔTK1761-TK1762-TK1763 This study
    TS559 TS517 with ΔTK0149c This study
    TS561 TS517 with ΔTK0148-TK0149 This study
    PdaD KU216 ΔpyrF with ΔTK0148-pdaD 6
Plasmids
    pTS502 pUC118::PhmtB-(His)10TK1827-trpE-TK1828 This study
    pTS503 pUC118::PhmtB-TK1827-trpE-TK1828 with TAG at codon 5 of TK1827 This study
    pTS517 pUC118::TK0662-TK0663-TK0664-trpE-TK0662-TK0663-TK0665 This study
    pTS521 pUC118::TK0662-TK0663-trpE-TK0665 This study
    pTS535 pUC118 with MCS1-(trp-6MPs)-MCS2d This study; Fig. S1
    pTS538 pTS535::TK1760-TK1764-(trp-6MPS)-TK1761 This study; Fig. S2
    pTS541 pTS535::TK1825-TK1826-TK1828-(trp-6MPs)-TK1827-TK1828 This study; Fig. S2
    pTS559 pTS535::TK0147-TK0148-TK0150-(trp-6MPs)-TK0150-TK0151 This study
    pTS561 pTS535::TK0147-TK0150-(trp-6MPs)-TK0150-TK0151 This study
    pLC70 trpE; Pgdh-PF1848e 19
    pTS436 pLC70::PhmtB-TK1761 This study
    pTS543 pLC70::PhmtB-TK0149 This study
a

pyrF = TK2276 (21); trpE = P2279-TK0254 (18); pdaD = TK0149 (6).

b

Loss of TK0664 results in resistance to 6-methyl purine (6MPr).

c

Loss of TK0149, or loss of TK0148 plus TK0149, results in agmatine auxotrophy.

d

The trp-6MPs cassette has P2279 and PhmtB directing the divergent transcription of trpE (TK0254) and TK0664, respectively. TK0664 expression results in sensitivity to 6-methyl purine (6MPs). Flanking the cassette are multiple cloning sites 1 and 2 (MCS1 and MCS2) that have single cleavage sites for HindIII, SphI, BglII, BclI, ClaI, NheI, SpeI, NotI, SacII, ApaI, AccI, AscI, EcoRV, and BstEII and for EcoRI, KpnI, Acc65I, BamHI, XbaI, respectively (see Fig. S1 in the supplemental material).

e

Expression of PF1848 confers resistance to simvastatin and mevinolin (15, 19).

f

Figures S1 and S2 may be found in the supplemental material.