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通过在盘基网柄菌中进行反义转化对盘基网柄菌I缺失突变体进行拟表型模拟。

Phenocopy of discoidin I-minus mutants by antisense transformation in Dictyostelium.

作者信息

Crowley T E, Nellen W, Gomer R H, Firtel R A

出版信息

Cell. 1985 Dec;43(3 Pt 2):633-41. doi: 10.1016/0092-8674(85)90235-1.

Abstract

Using an antisense construct of the discoidin gene transfected into Dictyostelium, we have repressed the expression of the three endogenous discoidin genes. Transformants exhibit a greater than 90% reduction in accumulated discoidin mRNA and protein. Nuclear run-on assays show that both the endogenous and the antisense genes are transcribed. Since only minor amounts of endogenous gene transcripts and none from the antisense gene can be detected on blots, we suggest that hybrids are formed within the nucleus and are rapidly degraded. Discoidin is believed to play a role in cell-substratum interaction and exhibits homologies to fibronectin. Discoidin-minus mutants exhibit the developmental phenotype of not streaming on a plastic surface. Antisense transformants show a similar phenotype and are thus phenocopies of these mutants.

摘要

通过将盘状球蛋白基因的反义构建体转染到盘基网柄菌中,我们抑制了三个内源性盘状球蛋白基因的表达。转化体中积累的盘状球蛋白mRNA和蛋白质减少了90%以上。核转录分析表明内源性基因和反义基因均被转录。由于在印迹上仅能检测到少量内源性基因转录本,而未检测到反义基因的转录本,我们推测在细胞核内形成了杂种并迅速降解。盘状球蛋白被认为在细胞与底物的相互作用中起作用,并且与纤连蛋白具有同源性。缺乏盘状球蛋白的突变体表现出在塑料表面不流动的发育表型。反义转化体表现出类似的表型,因此是这些突变体的拟表型。

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