Integration of a 2A-tagRFP reporter gene into tyr.
PCR amplification and sequence of 5’ junction fragments between tyr exon 4 and the targeted 2A-tagRFP-CAAX-Sv40 vector from randomly selected RFP-negative F0 injected embryos. 5/6 RFP-negative F0 injected embryos contain the expected 5’ junction fragment (marked with an ‘*’). The junction fragments from embryos F0-1 and −2 were TA-cloned and sequenced. 3 out of 4 cloned PCR products from embryo F0-1 and 3 out of 3 cloned products from embryo 2 showed precise 5’ integration in tyr. 1 of the 4 embryos from F0-1 contained a single nucleotide polymorphism in the 2A peptide sequence (shown in red).
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