Roshina M P, Loginova N N, Devin A B, Gvozdev V A
Genetika. 2008 Jun;44(6):752-60.
The 1.25-kb heterochromatic Stellate repeats of Drosophila melanogaster are capable of stably persisting in transgenic constructs and silencing the white reporter gene (mosaic position effect variegation). This system reveals an unusual form of silencing, which is insensitive to known modifiers of position effect variegation. The unusual form of silencing was studied with yeast Saccharomyces cerevisiae, a simple eukaryotic model. To be transferred into yeast cells, the D. melanogaster Stellate repeats were cloned in the pYAC4 centromeric vector (CEN4, URA3, TRP1, HIS3). The HIS3 and/or URA3 genes could be inactive in plasmids consisting of pYAC4 and the Stellate insert in yeast cells. Deletion of D. melanogaster DNA from the plasmid was found to activate the URA3 and HIS3 genes. It was assumed that the genes were repressed rather than damaged in the presence of the Stellate repeats and that a new form of gene silencing was revealed in S. cerevisiae.
黑腹果蝇1.25千碱基对的异染色质星状重复序列能够稳定存在于转基因构建体中,并使白色报告基因沉默(镶嵌位置效应斑驳)。该系统揭示了一种不同寻常的沉默形式,其对已知的位置效应斑驳修饰因子不敏感。利用简单的真核模型酿酒酵母对这种不同寻常的沉默形式进行了研究。为了转入酵母细胞,将黑腹果蝇星状重复序列克隆到pYAC4着丝粒载体(CEN4、URA3、TRP1、HIS3)中。在由pYAC4和酵母细胞中的星状插入片段组成的质粒中,HIS3和/或URA3基因可能失活。发现从质粒中删除黑腹果蝇DNA会激活URA3和HIS3基因。据推测,在存在星状重复序列的情况下,这些基因是被抑制而非受损,并且在酿酒酵母中揭示了一种新的基因沉默形式。