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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: J Microbiol Methods. 2011 Apr 29;86(2):156–159. doi: 10.1016/j.mimet.2011.04.014

Figure 1.

Figure 1

Opposing promoter constructs mediate RNAi. A. Schematic view of an RNAi plasmid with major elements shown; see text for details. TEL, telomeres; T, terminator; P, promoter; Target, sequence to be silenced. Restriction sites are indicated by vertical lines labeled with the abbreviated enzyme name. B. An opposing PGAL7 construct mediates silencing of ADE2 and LAC1. Left, A ura5 mutant (JEC43) was transformed with plasmid pIBB1, shown schematically, and grown on galactose medium lacking uracil (-ura/gal) for 2 days at 30°C. Right, a parallel study with plasmid pIBB29, showing a master plate with a random selection of transformants streaked onto niger seed agar with galactose (NSA/gal) and grown for 2 days at 30 °C. Controls streaked at the bottom of the plate are a wild-type strain that melanizes and appears brown (LAC1), and a deletion strain that does not melanize and therefore remains white (lac1).