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Figure 1


Fig. 1. Production of Zp3-Gja1 transgenic mice. (A) Structure of the Zp3-Gja1 transgene (not to scale). The transgene consisted of a 6 kb fragment of the Zp3 promoter, a 1.3 kb Gja1 exon 2 cDNA containing the entire rat Cx43 coding sequence (thin line), and a cassette consisting of a MYC-tag-encoding sequence and polyadenylation signal. The solid black section of the cDNA indicates the probe for Southern blotting and arrowheads represent the primers for PCR genotyping and RT-PCR. (B-D) Functional testing of Cx43-MYC. (B) Lucifer yellow dye was injected into a single cell (asterisk) of a population of gap-junctional-communication-deficient HeLa cells expressing the transgene. The dye readily passed to neighboring cells, as revealed by fluorescence microscopy (image at right). (C) The typical result of no dye transfer after injecting Lucifer yellow into un-infected HeLa cells not expressing the transgene. (D) Dye was injected into HeLa cells expressing native (not MYC-tagged) Cx43. (E) A typical Southern blot showing detection of the transgene in a sample of offspring. Digestion of tail-snip DNA with BanI generated a 1.3 kb fragment specific to the transgene in addition to the endogenous genomic fragment at 4.3 kb. Lanes 1-7 indicate results from seven of the potential founder offspring tested; lanes 5 and 7 represent transgenic lines 2b and 4, respectively, which were analyzed in the present study. Bar, 50 µm.





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