Function and anatomy of plant siRNA pools derived from hairpin transgenes-7

<p><b>Copyright information:</b></p><p>Taken from "Function and anatomy of plant siRNA pools derived from hairpin transgenes"</p><p>http://www.plantmethods.com/content/3/1/13</p><p>Plant Methods 2007;3():13-13.</p><p>Published online 25 Nov 2007</p><p>PMCID:PMC2217544.</p><p></p> mC9 sense and anti-sense sequences separated by the TGA1 intron (I) with expression by the CMV 35S promoter (35S) and NOS terminator. Transgene integrity for plants Tc9 #1, 3, 5, 9, 11, 13 was verified via PCR (primer position indicated by black arrows) to detect sense (s) and anti-sense (as) transgene elements. Endogenous rbcL is control. For siRNA expression 30 μg of total RNA from wt tobacco or Tc9 transformants was analysed by Northern blotting. A 31 nt mC9 DNA oligo (m) is size marker and an overnight phosphor-image is shown. In vivo gene silencing in Tc9 transformants. Transient transformation was performed by co-infiltration of tobacco wt, Tc9 #1 and #3 with plasmids expressing mC9 sense (pBmC9s) and EYFP transcripts (pBEYFP). Transcript levels in leaf regions were analysed 48 hr post-infiltration by Northern blotting (as described in materials and methods)