Figure 1From: Function and anatomy of plant siRNA pools derived from hairpin transgenesGeneration of transgenic tobacco expressing mC9 siRNAs. (A) mC9 transgene and siRNA expression. The mC9 siRNA cassette contains 400 bp 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. (B) 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).Back to article page