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. 2004 Dec 2;32(21):e171. doi: 10.1093/nar/gnh170

Figure 1.

Figure 1

Preparation of human H1 RNA gene promoter-based siRNA expression constructs. The 19 nt GUS gene specific sequence (GT1 or GT2) separated by a 9 nt spacer from the reverse complement of the same sequence followed by a termination signal of five thymidines (H1-GT1 or H1-GT2) was cloned into pSUPER (34) downstream of the H1 promoter (H1-P). The H1-P::GT expression construct harboring H1-GT1 or H1-GT2 was then excised and cloned into the binary vector pGPTV-HPT (41). The resulting vector, pGPH1-HPT-GT1 or pGPH1-HPT-GT2, which contained a hygromycin phosphotransferase (hpt) selectable marker gene under the control of a nopaline synthase promoter (Pnos)-transcription terminator (pAg7, agropine synthase polyadenylation signal sequence) pair, was then mobilized into A.tumefaciens C58 for transforming tobacco. The predicted secondary siRNA structures of H1-GT1 and H1-GT2 are depicted.