Ahmad K, Golic K G
Department of Biology, University of Utah, Salt Lake City 84112, USA.
Genetics. 1996 Oct;144(2):657-70. doi: 10.1093/genetics/144.2.657.
A transgene was inserted at several different chromosomal sites in Drosophila melanogaster, where its expression was subject to genomic position effects. Quantitative position effects and variegated and constant patterned position effects were observed. We investigated the status of the affected gene in the somatic cells where it normally functions. The FLP site-specific recombinase was used to remove the gene from the chromosome and its expression was then evaluated. We show that the FLP recombinase functions in cells that have finished their developmental program of mitoses. When FLP acts on directly repeated copies of its target site (FRT), the DNA flanked by those FRTs is excised from the chromosome as a closed circle. The extrachromosomal circle is maintained in nondividing cells, and a gene located on such a circle can be expressed. We then demonstrate that a gene subject to either variegated or constant position effect can be relieved of that effect by excision of the gene from the chromosome in cells where it would otherwise be inactive. We also observed a strong inhibition of FLP-mediated recombination for target sites located near centric heterochromatin.
一个转基因被插入到黑腹果蝇的几个不同染色体位点,其表达受到基因组位置效应的影响。观察到了定量位置效应以及斑驳和恒定模式的位置效应。我们研究了该受影响基因在其正常发挥功能的体细胞中的状态。使用FLP位点特异性重组酶从染色体上移除该基因,然后评估其表达。我们表明,FLP重组酶在已经完成有丝分裂发育程序的细胞中起作用。当FLP作用于其靶位点(FRT)的直接重复拷贝时,位于这些FRT两侧的DNA作为一个闭环从染色体上切除。染色体外的环在不分裂的细胞中得以维持,位于这样一个环上的基因可以表达。然后我们证明,一个受到斑驳或恒定位置效应影响的基因,可以通过在否则会无活性的细胞中将该基因从染色体上切除而解除这种效应。我们还观察到,对于位于着丝粒异染色质附近的靶位点,FLP介导的重组受到强烈抑制。