Skip to main content
. 2007 Dec;145(4):1272–1281. doi: 10.1104/pp.107.106062

Figure 1.

Figure 1.

Structural features of the pSAT-RNAi vector system. A, The general structure of a pSAT-RNAi plasmid and its cloning compatibility with pRCS2-based binary plasmid. A typical pSAT-RNAi plasmid is composed of a promoter and terminator region, the ChsA intron, and two unique multicloning sites, MCS-I and MCS-II, which should be used for cloning the two target gene sequences in reverse orientations. The entire hpRNA cassette is flanked by AgeI and NotI, allowing its transfer between different pSAT plasmid backbones. The hpRNA cassette is also flanked with rare cutters (e.g. PI-PspI), which can be used for its cloning into the pRCS2-based binary plasmid. B, Outline of the different pSAT-RNAi plasmids produced in this study. Expression of hpRNA is controlled by various promoters and terminators and each of the various RNAi cassettes can be transferred to the pRCS2-based binary plasmid using different combinations of rare-cutting endonucleases, as indicated. Both MCS-I and MCS-II are present in all plasmids and restriction recognition sites that are not unique in all plasmids are indicated by asterisks. Plasmid names from top to bottom are: pSAT3.masP.RNAi, pSAT4.35SP.RNAi, pSAT5.nosP.RNAi, pSAT6.masP.RNAi, pSAT6.35SP.RNAi, pSAT6.supP.RNAi, and pSAT6.rbcP.RNAi. 35S, CaMV 35S promoter and terminator; sup, superpromoter; mas, manopine synthase promoter and terminator; rbc1, Rubisco small-subunit promoter and terminator; nos, nopaline synthase promoter and terminator; ags, agropine synthase terminator. One, two, and three asterisks represent BspEI, BamHI, and XbaI restriction sites, respectively.