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. 2013 Oct 24;8(10):e78119. doi: 10.1371/journal.pone.0078119

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© 2013 Sano et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

PMC Copyright notice

DT40 clones carrying modified hChr14 (CH2D4) and modified hChr2/hChr22 (SLKD18), respectively, were fused by WCF to generate DT40 hybrid clone cKSLD22. The presence of the two modified human chromosomes in cKSLDH22 clone was confirmed by FISH with human COT-1 DNA as the probe. Transfection of cKSLDH22 clone with Cre-expression plasmid induced for site-specific translocation between the two loxP sites at the cos138 locus on the SLK HAC and another at the RNR2 locus on the CH2D HAC. The presence of the three human immunoglobulin loci, hIgH, hIgK and hIgL, as a single HAC was confirmed by three-color FISH, with the DNA probes for hIgH, hIgK and hIgL labeled with SpectrumRed, SpectrumGreen, and SpectrumOrange, respectively.