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14
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0042042035
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unpublished results
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The genes encM and encK are present on different transcripts. Their respective downstream genes encN and encJ are also inactivated as a result of polar effects in each mutant. Inactivation of these starter unit biosynthesis genes resulted in a 0% and 25% reduction in 1 biosynthesis, respectively, suggesting that EncN is extraneous and that other cellular thiolases can complement the loss of the dedicated thiolase EncJ (Xiang, L.; Moore, B. S., unpublished results).
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Xiang, L.1
Moore, B.S.2
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15
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16
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0042042036
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note
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10 The encK single-crossover mutant S. maritimus KK was selected after propagating transconjugants on SGGP plates at 37°C. and apramycin-resistant colonies were confirmed by Southern hybridization with biotinylated encK. Biotin-labeling and detection of chemiluminescent positives were performed with the DNADetector HPR Southern Blotting Kit (KPL. Inc.) according to the manufacturer. In a similar fashion, the encM disruption vector pBM1 was constructed from an 879 bp internal fragment of encM amplified with the primers 5′-CGGACACAGCATGGCCGGGC-3′ and 5′-GTCGTCGGGCACGCGCGCC-3′ and conjugated into "S. maritimus" to yield the encM single-crossover mutant "S. maritimus" KM.
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17
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0042542862
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note
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2O to 0.085% TFA in MeCN over a period of 40 min.
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18
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0041540773
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note
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18 (Waters, 20 mm × 250 mm, 10 μm) HPLC employing gradient elution (40% MeOH/0.15% TEA to 100% MeOH over 50 min) at flow rate of 9.5 mL/min. The resulting fractions were pooled on the basis of retention time in the UV chromatogram yielding wailupemycin E (4, 5.9 mg), wailupemycin D (3, 3.4 mg), wailupemycin F (5, 5.4 mg), and wailupemycin G (6, 9.4 mg), which eluted at 17, 22, 27, and 28.5 min, respectively.
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19
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0043043721
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note
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6, Δ + 0.4 mmu).
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20
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0025310579
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Zhang, H. L.; He, X. G.; Adefarati, A.; Gallucci, J.; Cole, S. P.; Beale, J. M.; Keller, P. J.; Chang, C. J.; Floss, H. G. J. Org. Chem. 1990, 55, 1682-1684.
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Zhang, H.L.1
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Cole, S.P.5
Beale, J.M.6
Keller, P.J.7
Chang, C.J.8
Floss, H.G.9
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21
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0043043722
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note
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6, Δ -0.1 mmu).
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22
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0042542825
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note
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5, Δ + 0.6 mmu).
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23
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0027970295
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McDaniel, R.; Ebert-Khosla, S.; Fu, H.; Hopwood, D. A.; Khosla, C. Proc. Natl. Acad. Sci. U.S.A. 1994, 91, 11542-11546.
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24
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0042542833
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Harris, T. M.; Harris, C. M. Pure Appl. Chem. 1986, 58, 283-294. Shen, Y.; Yoon, P.; Yu, T.-W.; Floss, H. G.; Hopwood, D. A.; Moore, B. S. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 3622-3627.
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Harris, T.M.1
Harris, C.M.2
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25
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Harris, T. M.; Harris, C. M. Pure Appl. Chem. 1986, 58, 283-294. Shen, Y.; Yoon, P.; Yu, T.-W.; Floss, H. G.; Hopwood, D. A.; Moore, B. S. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 3622-3627.
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Moore, B.S.6
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26
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0029670331
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Alvarez, M. A.; Fu, H.; Khosla, C.; Hopwood, D. A.; Bailey, J. E. Nat. Biotechnol. 1996, 14, 335-338.
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