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. 2006 Nov 14;5:33. doi: 10.1186/1475-2859-5-33

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Copyright © 2006 Steinborn et al; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Transformation procedure based on simultaneous integration of the plasmids pAL-HPH-phbA-phbB and pAL-ALEU2m-phbC into the 25S rDNA of A. adeninivorans G1211 (aleu2). The two plasmids pAL-HPH-phbA-phbB and pAL-ALEU2m-phbC containing the expression cassettes with phbA, phbB and phbC genes are linearised by BglII or Esp3I digestion, respectively. The resulting fragments flanked by 25S rDNA sequences are co-integrated into the 25S rDNA by homologous recombination. Transformants are selected either by resistance to hygromycin B (plasmid pAL-HPH-phbA-phbB [25,44]) or the complementation of the aleu2 mutation (plasmid pAL-ALEU2m-phbC [20]).