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. 2007 Jun 27;2007:91409. doi: 10.1155/2007/91409

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Copyright © 2007 Samiha Sioud et al.

This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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(a) Southern blot analysis confirming insertion of pSS3 plasmid into the Streptomyces sp. US24 chromosome via homologous recombination. Blots were probed with the 696 bp DNA fragment (right panel) or the whole linearized pSS2 (left panel). Lanes 4 and 8, 1 kb ladder used as molecular marker, lanes 2 and 6, wild-type genomic DNA cut with BamHI, lanes 1 and 5, apramycin resistant (Apr^R) exconjugant mutant genomic DNA cut with BamHI, and lanes 3 and 7, the used probes linearized pSS2 and 696 bp DNA fragment, respectively. (b) Schematic representation of the integration of plasmid pSS3 in the chromosome of Streptomyces sp. US24 by homologous recombination. The resident copy of the 696 bp locus involved in the biosynthetic pathway of Cyclo (L-Phe, L-Pro) DKP of the US24 strain (shaded boxes) present in the chromosome of Streptomyces sp. US24 is shown. del int: deletion of a part of the integrase-encoding gene by partial digestion HindIII; aac(3)IV: apramycin resistance gene; B: restriction site for BamHI; E: restriction site for EcoRI.

(a) Southern blot analysis confirming insertion of pSS3 plasmid into the Streptomyces sp. US24 chromosome via homologous recombination. Blots were probed with the 696 bp DNA fragment (right panel) or the whole linearized pSS2 (left panel). Lanes 4 and 8, 1 kb ladder used as molecular marker, lanes 2 and 6, wild-type genomic DNA cut with BamHI, lanes 1 and 5, apramycin resistant (Apr^R) exconjugant mutant genomic DNA cut with BamHI, and lanes 3 and 7, the used probes linearized pSS2 and 696 bp DNA fragment, respectively. (b) Schematic representation of the integration of plasmid pSS3 in the chromosome of Streptomyces sp. US24 by homologous recombination. The resident copy of the 696 bp locus involved in the biosynthetic pathway of Cyclo (L-Phe, L-Pro) DKP of the US24 strain (shaded boxes) present in the chromosome of Streptomyces sp. US24 is shown. del int: deletion of a part of the integrase-encoding gene by partial digestion HindIII; aac(3)IV: apramycin resistance gene; B: restriction site for BamHI; E: restriction site for EcoRI.