Roseman R R, Johnson E A, Rodesch C K, Bjerke M, Nagoshi R N, Geyer P K
Department of Biochemistry, University of Iowa, Iowa City 52242, USA.
Genetics. 1995 Nov;141(3):1061-74. doi: 10.1093/genetics/141.3.1061.
P elements are widely used as insertional mutagens to tag genes, facilitating molecular cloning and analyses. We modified a P element so that it carried two copies of the suppressor of Hairy-wing [su(Hw)] binding regions isolated from the gypsy transposable element. This transposon was mobilized, and the genetic consequences of its insertion were analyzed. Gene expression can be altered by the su(Hw) protein as a result of blocking the interaction between enhancer/silencer elements and their promoter. These effects can occur over long distances and are general. Therefore, a composite transposon (SUPor-P for suppressor-P element) combines the mutagenic efficacy of the gypsy element with the controllable transposition of P elements. We show that, compared to standard P elements, this composite transposon causes an expanded repertoire of mutations and produces alleles that are suppressed by su(Hw) mutations. The large number of heterochromatic insertions obtained is unusual compared to other insertional mutagenesis procedures, indicating that the SUPor-P transposon may be useful for studying the structural and functional properties of heterochromatin.
P 元件被广泛用作插入诱变剂来标记基因,以促进分子克隆和分析。我们对一个 P 元件进行了改造,使其携带从吉普赛转座元件中分离出的两个毛翅抑制因子[su(Hw)]结合区域的拷贝。这个转座子被激活,并分析了其插入的遗传后果。由于 su(Hw)蛋白阻断了增强子/沉默子元件与其启动子之间的相互作用,基因表达可能会发生改变。这些效应可以在远距离发生且具有普遍性。因此,一个复合转座子(抑制因子-P 元件,即 SUPor-P)将吉普赛元件的诱变效力与 P 元件可控制的转座作用结合起来。我们表明,与标准 P 元件相比,这个复合转座子会导致突变类型的增加,并产生可被 su(Hw)突变抑制的等位基因。与其他插入诱变程序相比,获得的大量异染色质插入是不同寻常的,这表明 SUPor-P 转座子可能有助于研究异染色质的结构和功能特性。