Figure 2.

TALEN-Mediated Mutagenesis in M. truncatula Using Direct Cloning Vectors.

(A) Maps of two M. truncatula genes targeted for mutagenesis using a single TALEN pair. PCR primers used for screening in (B) are shown as green arrowheads. The sequence of the TALEN target site is shown with TALEN binding sites underlined.

(B) HaeIII PCR digestion screening of eight T1 progeny of plant WPT52-4. The amplified locus is indicated on the right. bar and ACTIN are controls that amplify the transgene and a native gene, respectively. HM340 (D), wild-type product digested with HaeIII; HM340 (UD), undigested wild-type product.

(C) DNA sequences of undigested amplicons from plant WPT52-4-4. The reference sequence of the unmodified locus is shown on the top. TALEN binding sites are underlined. HaeIII restriction site used for the screening is in red.