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. 2004 May 10;101(20):7833–7838. doi: 10.1073/pnas.0402267101

Fig. 1.

Fig. 1.

Generation of transgenic Arabidopsis expressing Dof1. (a) The metabolic pathway for nitrogen assimilation in plants. PEP, phosphoenolpyruvate; OAA, oxaloacetate; GOGAT, glutamate synthase; NIA, nitrate reductase. (b) Construct of the plasmid containing a derivative of the CaMV 35S promoter (35S), Dof1 cDNA fused to the sequence for an epitope tag (HA), and the nopaline synthase terminator (nos) between the right (RB) and left (LB) borders of the T-DNA. The kanamycin-resistance gene was located between the RB and the derivative of the 35S promoter. (c) RT-PCR analysis of expression of the Dof1 transgene. Total RNA was prepared from the vector control plants (lane C) or the transgenic Dof1 lines (lanes 1–3). PCR primers specific to the Dof1 transgene or β-tubulin gene were used. (d) Immunological detection of Dof1 with anti-HA antibodies. Nuclear protein from the same amount of leaf tissue was loaded on each lane. Loading a similar amount of nuclear protein was verified with antihistone H1 antibodies. (e) Photographs of the control and Dof1 transgenic plants. The plants were grown on the plates for 2 weeks and further grown in soil.