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MOESM4 of Recapitulation of gametic DNA methylation and its post-fertilization maintenance with reassembled DNA elements at the mouse Igf2/H19 locus

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posted on 15.01.2020 by Hitomi Matsuzaki, Daichi Kuramochi, Eiichi Okamura, Katsuhiko Hirakawa, Aki Ushiki, Keiji Tanimoto
Additional file 4: Figure S4. Generation and structural analysis of YAC-TgM carrying the LCb and LCb118 fragments. (A) Structure of the 150-kb human β-globin locus YAC. The LCR and β-like globin genes are denoted as gray and filled boxes, respectively. The enlarged map shows tandemly arrayed LCb and LCb118 fragments, inserted 3′ to the LCR for employing co-placement strategy. The positions of loxP5171 and loxP2272 are indicated as solid and open triangles, respectively. The expected SfiI restriction enzyme fragments (thick lines) and probes (filled rectangles) used in (B) are shown. (B) Long range structural analysis of the LCb-LCb118 YAC transgene. DNA from thymus cells was digested with SfiI in agarose plugs and separated by pulsed-field gel electrophoresis, and Southern blots were hybridized separately to probes. (C) In vivo Cre-loxP recombination to derive LCb or LCb118 TgM. Tail DNA from parental and daughter YAC-TgM sublines was digested with KpnI and analyzed by Southern blotting using the probe.


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