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. 2017 Aug 4;40(4):1067–1077. doi: 10.3892/ijmm.2017.3091

Figure 1.

Figure 1

Development of breast cancer associated gene 3 (BCA3) transgenic mice with targeted overexpression of murine BCA3 in mature osteoclasts. (A) The targeting vector included 23 bp from the 5′ end of cathepsin K (Ctsk) exon 1, the entire BCA3 cDNA sequence (including the endogenous stop codon), a neomycin selection cassette flanked by frt sites and 13 base pairs from the 3′ end of exon 2. This construct was integrated into the Ctsk gene exon 1 within the BAC clone by homologous recombination. (B) Upper panel: after selecting positive clones, the Neo cassette was excised by inducing FLP recombinase F70L in the DH10B bacterial host of the recombined BAC clone. The blue and red large horizontal arrows show the location of the genotyping primers; Lower panel: agarose gel of PCR amplicons demonstrating the expected 543 bp fragment from the transgene. (C) The tibiae and femurs were isolated from mice from two different transgenic lines and RNA extracted. Real-time PCR demonstrated that both lines overexpress BCA3. Data were analyzed by unpaired t-test. (**p<0.01).