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. Author manuscript; available in PMC: 2010 Mar 15.
Published in final edited form as: J Immunol. 2009 Mar 15;182(6):3866–3876. doi: 10.4049/jimmunol.0713949

Fig. 1. Generation of Epilysin-null (Mmp28−/−) Mice.

Fig. 1

A, The targeting construct was designed to replace most of the exons, including exon 5, which codes for the catalytic domain. A diphtheria-toxin cassette was included to selected against ES cells with mistargeted recombination. The targeting construct was linearized with XhoI and injected in C57Bl/6 blastocyst. After recombination, exons 4 to 7 and the full coding portion of the exon 8 are removed, and the epilysin transcript only contains exons 1 to 3 (coding for the pro-region only). B, ES clones (144 total) were screened by Southern hybridization using a 694 bp probe in the 3'terminal end of intron 1 just outside of the targeting construct. With Bam HI digestion, wildtype epilysin gene gives a ∼9 kb band while targeted gene gives a ∼7.5 kb band. C, F1 offspring of chimeric mice were screened by Southern and PCR assays. For the PCR assay, the recombined locus produces a larger DNA fragment (324 nt) than that from the wildtype gene (259 nt) and both bands from heterozygotes. Three primers were used, the position and orientation of which are indicated by arrowheads: a wildtype forward primer, a neomycin cassette forward primer, and a common reverse primer.