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. Author manuscript; available in PMC: 2010 Jul 1.

Published in final edited form as: Dev Dyn. 2009 Jul;238(7):1727–1743. doi: 10.1002/dvdy.21994

Fig. 3 — Southern blot analysis of genomic DNA harvested from SB-mediated transgenic Xenopus tropicalis and Xenopus laevis. Genomic DNA harvested from progeny of each of the founder animals was digested with BglII and separated by electrophoresis on an agarose gel, transferred to a membrane and probed with a radiolabelled GFP-encoding DNA fragment. a: Schematic representation of the pT2βGFP SB transposon indicating the approximate position of the unique BglII site and region used for the probe (bar). Not to scale. b: Southern blot analysis of pT2βGFP transgenic founders. Outcross of founder 4M resulted in two discrete hybridization patterns indicating independent segregation of the alleles (compare samples 4M-1 and 4M-2). The founder animal contains, at least, four copies of the GFP sequence that are inherited by the progeny (open and closed triangles). Likewise, the progeny of 8F display two different hybridization patterns (compare 8F-1 and 8F-3). Founder 8F also contains at least four copies of the transgene (^ and *s). Progeny from founder 7M have a complex hybridization pattern suggesting the presence of a concatamer of transposon transgenes. Tadpoles 4M-1 and 7M have hybridizing bands (labeled open triangle and #) that migrate faster than the predicted lower limit for the BglII digested transgene (2.97 kb; see Fig. 1a). This indicates that the integration events at these loci are complex and have involved fragmentation of the transposon transgene. Size markers (in kb) are indicated on the left side of the blot. c: Enlarged view of the Southern blot to illustrate that founder 6M has two closely migrating bands (*). d: Southern blot analysis of Xenopus laevis founder lines L2M, L3M and L6M. Genomic DNA samples for three GFP positive F₁ animals (#1, 2 and 3) and a GFP negative F₁ sibling (#4) from each founder line were digested with BglII, separated on a 0.7% (w/v) agarose gel, and probed with a 700 bp GFP fragment as in figure 3b. L2M founder line has at least 5 hybridizing GFP bands, L3M has at least 3 GFP positive bands and L6M has two GFP positive bands.

Fig. 3 — Southern blot analysis of genomic DNA harvested from SB-mediated transgenic Xenopus tropicalis and Xenopus laevis. Genomic DNA harvested from progeny of each of the founder animals was digested with BglII and separated by electrophoresis on an agarose gel, transferred to a membrane and probed with a radiolabelled GFP-encoding DNA fragment. a: Schematic representation of the pT2βGFP SB transposon indicating the approximate position of the unique BglII site and region used for the probe (bar). Not to scale. b: Southern blot analysis of pT2βGFP transgenic founders. Outcross of founder 4M resulted in two discrete hybridization patterns indicating independent segregation of the alleles (compare samples 4M-1 and 4M-2). The founder animal contains, at least, four copies of the GFP sequence that are inherited by the progeny (open and closed triangles). Likewise, the progeny of 8F display two different hybridization patterns (compare 8F-1 and 8F-3). Founder 8F also contains at least four copies of the transgene (^ and *s). Progeny from founder 7M have a complex hybridization pattern suggesting the presence of a concatamer of transposon transgenes. Tadpoles 4M-1 and 7M have hybridizing bands (labeled open triangle and #) that migrate faster than the predicted lower limit for the BglII digested transgene (2.97 kb; see Fig. 1a). This indicates that the integration events at these loci are complex and have involved fragmentation of the transposon transgene. Size markers (in kb) are indicated on the left side of the blot. c: Enlarged view of the Southern blot to illustrate that founder 6M has two closely migrating bands (*). d: Southern blot analysis of Xenopus laevis founder lines L2M, L3M and L6M. Genomic DNA samples for three GFP positive F₁ animals (#1, 2 and 3) and a GFP negative F₁ sibling (#4) from each founder line were digested with BglII, separated on a 0.7% (w/v) agarose gel, and probed with a 700 bp GFP fragment as in figure 3b. L2M founder line has at least 5 hybridizing GFP bands, L3M has at least 3 GFP positive bands and L6M has two GFP positive bands.