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Figure 1 | BMC Biotechnology

Figure 1

From: Efficient generation of long-distance conditional alleles using recombineering and a dual selection strategy in replicate plates

Figure 1

Generation of the targeting vector. (A) The retrieval vector was constructed by cloning Sal I/EcoR I and EcoR I/BamH I digested homology arms (colored boxes; about 500 bp) that had been generated by PCR amplification of indicated segments from BAC DNA into BamH I/Sal I digested pBR322. The DTA-SV40pA cassette was inserted into the Rsr II site of 3'RV-2. All four exons (open boxes) of il-13 and the last two exons of il-4 are shown. (B) The EcoR I-linearized retrieval vector was used to retrieve a 27 kB segment from the BAC by gap-repair in DY380 cells. The final targeting vector was generated by insertion of two mini targeting vectors containing either a blasticidin resistance cassette (5'MTV) or a neomycin/kanamycin resistance cassette (3'MTV) flanked by 350–500 bp of homology arms into the second intron of il-13 and downstream of il-4 by recombineering in DY380 cells.

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BMC Biotechnology

ISSN: 1472-6750

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