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. 2012 Nov 1;161(1):36–47. doi: 10.1104/pp.112.205369

Figure 1.

Figure 1.

A, Diagram of plasmid pSH-Tnt1 containing the Tnt1 element in the binary vector pZY101. LB and RB, Left and right borders, respectively; bar, gene conferring glufosinate resistance; 35S, promoter 35S; LTR, long terminal repeat. NdeI restriction sites and PCR fragments for bar (barA-B) and Tnt1 (TntA-B, TntC-D and TntE-F) amplification are shown. PCR fragment barA-B or TntC-D was used in Southern-blot analysis to probe for bar or Tnt1, respectively. B, Southern-blot analysis of Tnt1 primary transgenic lines to identify Tnt1-hybridizing bands. Chromosomal DNA (15–20 µg) from each transgenic line was digested with NdeI and probed with the TntC-D PCR fragment. The arrow indicates hybridization bands representing nontransposed Tnt1 (i.e. T-DNA associated). M, Molecular weight markers. Lanes 1 to 16 show Tnt1 mutants BS2-5, BS2-6, BS3-5, BS5-4, BS5-6, BS5-10, BS5-12, BS5-13, BS6-19, BS6-20, BS7-5, BS7-7, BS7-8, BS7-10, BS8-5, and BS8-7, respectively. cv Maverick is the parent line. C, Southern-blot analysis of Tnt1 transgenic lines to identify bar-hybridizing bands. The blot used in B was stripped and rehybridized using the barA-B PCR fragment as a probe.