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. 2002 Feb;13(2):445–453. doi: 10.1091/mbc.01-04-0211

Figure 1.

Figure 1

Constructs used for β-gal gene silencing and their efficiency. In the first two lanes, the construct introduced into the cells is described. In the third lane, the average β-gal activity and SD of randomly chosen tested clones are given in units per milligram of total protein. The last lane shows how many clones have been tested and how many of them exhibited gene silencing. As (a) a control an empty vector was introduced; (b) an antisense copy (800-bp fragment) of the transgene was introduced (V18 promoter), (c) an additional copy (800-bp fragment) of the transgene controlled by the V18 promoter was introduced (equivalent to cosuppression approaches in plants), (d) sense and antisense RNA both under the control of the V18 promoter were expressed from different gene constructs in the same vector, (e) the 800-bp gene fragment was inserted between two opposing promoters (V18 and actin15), (f) an 800-bp inverted repeat with a 500-bp linker, corresponding to β-gal gene sequences, was fused to the V18 promoter, and (g) an 800-bp fragment was inserted between two T7 promoters and transformed bacteria (strain BL21) were used to feed Dictyostelium cells. Only in e was silencing of the transgene observed.

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