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. 2009 May 14;4(5):e5553. doi: 10.1371/journal.pone.0005553

Figure 4. Shuffling of trypsinogen.

Figure 4

(A) Alignment of the aminoacid sequence of bovine cationic trypsinogen (BC), bovine anionic trypsinogen (BA) and human cationic trypsinogen (HC). Nucleotide sequence of the chosen recombination sites is shown. (B) Map of the 27 trypsinogen module plasmids, the acceptor vector, and of an example of one of the resulting shuffled construct obtained. B, BsaI restriction site. S, K: spectinomycin and kanamycin resistance genes. RB/LB, T-DNA right and left borders. AttB, Phage C31 recombination site, N tobamoviral 3′ non-translated region, T, Nos terminator. (C) Ethidium bromide-stained gels of 28 minipreps prepared from single colonies (1 to 24) or from 4 libraries (L1–4, approximately 700 clones in each) digested with XmaI (incorrect pattern 1, 2, 7 and 17).