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. Author manuscript; available in PMC: 2011 Jul 14.
Published in final edited form as: Chembiochem. 2007 Apr 16;8(6):632–641. doi: 10.1002/cbic.200600528

Figure 1.

Figure 1

Inactivation of valC in S. hygroscopicus 5008. A) Schematic representation of the replacement of a 913 bp internal fragment of valC with the 1.4 kb aac(3)IV. In shuttle plasmid pJTU474, aac(3)IV was inserted between 1.54 kb and 1.36 kb genomic fragments originally flanking the 913 bp region. While wild-type 5008 should give a 1.7 kb PCR-amplified product, mutant ZYR-1 should yield a 2.3 kb product using a pair of primers (ValC-F and ValC-R); B) PCR analysis of wild-type 5008 and mutant ZYR-1. PCR products were run on an agarose gel; C) HPLC comparison between cultures of the wild-type strain 5008 and the mutant ZYR-1. The peak corresponding to validamycin A is absent in mutant ZYR-1 cultures; D) Bioassay of fermentation broths of wild-type strain (5008), valC mutant (ZYR-1) and valC complemented mutant (ZYR-1/pJTU704). The agar plugs in the center of each plate are the indicator strain, Pellicularia sasakii. The fermentation supernatant of each sample was mixed with melted agar. The valC mutant did not produce validamycin A but regained the wild-type capability of producing the antibiotic after complementation with the shuttle plasmid pJTU704 carrying the full-length valC.