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Figure 5 | Plant Methods

Figure 5

From: Protocol: Precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli

Figure 5

Replacement of AtSTM exon 1 by in-frame fusion of GAPc2ox1. A: Schematic representation of the strategy used to generate the GAPc2ox1 cassette from pNosTerFRT-neo (plus GAPc2ox1) plasmid and target it into the AtSTM locus (captured in pBlueAtSTM17) by in-frame substitution of exon 1 sequences. B: FIGE gel results of Pme I and EcoR V digested pSTM17::GAPc2ox1 from two independent recombination events (lanes 2 and 3), clearly showing the increased size of the linearised construct (Pme I digest) and the additional fragment (Eco RV digest) due to the recombination of the GAPc2ox1 cassette. Lane 1 shows the control result with the original pBlueAtSTM17. M1 = 1 kb ladder (New Endgland Biolabs); M2 = High Molecular Weight Marker (Invitrogen).

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Plant Methods

ISSN: 1746-4811