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. 2011 Aug;77(15):5505–5512. doi: 10.1128/AEM.02070-10

Fig. 4.

Fig. 4.

Gene replacement by double-crossover recombination in R. marinus. (A) Double-crossover recombination between the linear crtBI deletion molecule and the chromosome of R. marinus SB-62 (ΔtrpB ΔpurA). The known sequence of 3,950 bp surrounding the carotenoid biosynthesis genes crtB (partial, 814 bp) and crtI (1,515 bp) in the wild-type strain PRI 493 is shown schematically. ORF1 encodes a putative peptidase (378 bp). The trpB marker cassette (1.4 kb) containing the groESL upstream sequence (200 bp) replaced a 2.4-kb MluI fragment. MluI sites (M) are indicated. (B and C) Southern analysis of MluI-digested genomic DNA. Lanes M, HindIII-digested λ DNA as a size marker (kb). Lane 1, strain PRI 493 (wt); lane 2, SB-62 (ΔtrpB ΔpurA); lanes 3 and 4, strains SB-71 and SB-72 (ΔtrpB ΔpurA crtBI′::trpB). The membrane was probed with the deleted MluI fragment (B) and trpB (C). For detection of the size marker, labeled λ DNA was added with each probe.