FIGURE 1
FROM:
A complete collection of single-gene deletion mutants of Acinetobacter baylyi ADP1
Véronique de Berardinis, David Vallenet, Vanina Castelli, Marielle Besnard, Agnès Pinet, Corinne Cruaud, Sumitta Samair, Christophe Lechaplais, Gabor Gyapay, Céline Richez, Maxime Durot, Annett Kreimeyer, François Le Fèvre, Vincent Schächter, Valérie Pezo, Volker Döring, Claude Scarpelli, Claudine Médigue, Georges N Cohen, Philippe Marlière, Marcel Salanoubat & Jean Weissenbach
doi:10.1038/msb.2008.10
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Method of construction of the single-gene deletion mutants by creation of a spliced PCR integration cassette. P1–P6 are used for integration cassette construction and P7, P8, S1 and S2 for verifications. The kanR integration cassette is obtained by PCR amplification using P1 and P2 primers on pEVL186 DNA template. The flanking regions, specific for the target gene, are amplified on wild-type DNA template by P3/P4 primers (R1 region) and P5/P6 primers (R2 region). Designations followed by a prime (') represent reverse complement sequences. The primers P7 and P8 are used for external PCR verification of the correct replacement of the targeted gene by the integrative cassette. The primers S1 and S2 located within the kanR cassette are used to sequence junctions of the cassette on the P7/P8 PCR product.