Skip to main content
. Author manuscript; available in PMC: 2010 Oct 1.
Published in final edited form as: Genesis. 2009 Oct;47(10):680–687. doi: 10.1002/dvg.20547

Figure 1. Generation of Hoxc8 conditional loss-of-function and Hoxc8 replaced by Hoxc9 mouse alleles.

Figure 1

(a) Hoxc8 genomic locus. E1 and E2 represent Exon1 and 2, yellow boxes represent coding region, purple lines represent the intron, and pink boxes represent the 3´ untranslated region (UTR). X=Xba1, H=HindIII, Sp=Sph1, Sc=SacII, N=Nco1, Sl=Sal1, Ea=Ear1. Targeting constructs contain ~9.5kb genomic DNA from the 5´ Sph1 site to the 3´ Xba1 site. “IR” represents internal ribosome reentry site, “marker” represents either eGFP or nlsLacZ reporter used in various versions of the targeting constructs. A Black box indicates the HindII-Sph1 fragment used as a probe for the Southern blot shown in (b). Black triangles indicate the positions of LoxP1 and LoxP3 primers used in (c).

(b) Southern analyses. Genomic DNA digested with Xba1 and probed with a 500bp HindIII-Sph1 probe identified a ~10.4kb wild-type (+) band. For the Hoxc8-LacZ allele, the size of the targeted band (F) is ~19.2kb (group 1). For the Hoxc8-GFP, Hoxc8->c9-GFP, and Hoxc8->c9-LacZ alleles, the size of the targeted band (F) is ~7kb (group 2).

(c) PCR reactions using LoxP1 and LoxP3 primers. The wild-type allele (+) produces a ~230bp band while the targeted allele (F) produces a ~300bp band.