Scherczinger C A, Knecht D A
Department of Molecular and Cell Biology, University of Connecticut, Storrs.
Antisense Res Dev. 1993 Summer;3(2):207-17. doi: 10.1089/ard.1993.3.207.
We have previously demonstrated effective inhibition of Dictyostelium discoideum myosin heavy chain (MIIHC) gene expression after introduction of an antisense gene on a transformation vector into cells. We now report that transcription of a sense orientation MIIHC gene has a similar, though not as dramatic, effect on expression from the endogenous gene. Cells transformed with a sense fragment of the MIIHC gene are defective in cytokinesis and become giant and multinucleated when growing on a surface. This morphological phenotype is correlated with decreased expression of the endogenous MIIHC gene. Sense transformants accumulate less MIIHC protein and mRNA than control transformed cells, concomitant with accumulation of a vector-derived sense transcript. Expression of the introduced sequences seems to be required for the sense effect. The possible causes of this surprising phenomenon are discussed.
我们之前已经证明,将反义基因导入转化载体后,可有效抑制盘基网柄菌肌球蛋白重链(MIIHC)基因的表达。我们现在报告,有义方向的MIIHC基因转录对内源基因的表达有类似的影响,尽管不那么显著。用MIIHC基因的有义片段转化的细胞在胞质分裂方面存在缺陷,在表面生长时会变成巨大的多核细胞。这种形态学表型与内源MIIHC基因表达的降低相关。有义转化体比对照转化细胞积累的MIIHC蛋白和mRNA更少,同时积累了载体衍生的有义转录本。有义效应似乎需要导入序列的表达。我们讨论了这一惊人现象的可能原因。