Singh M, Silva E, Schulze S, Sinclair D A, Fitzpatrick K A, Honda B M
IMBB and Department of Biological Sciences, Simon Fraser University, Burnaby, Canada.
Gene. 2000 Apr 18;247(1-2):167-73. doi: 10.1016/s0378-1119(00)00102-5.
We report here on the cloning and characterization of a new theta-class glutathione-S-transferase (GST) gene, gst-3, from Drosophila melanogaster. Its sequence is distinct from previously characterized Drosophila GST genes, and Southern blotting shows no other closely related genes in the genome. In-situ hybridization localizes the gene to chromosome 2 (55D), near gst-2 (53F), and clearly separate from the gst-D cluster at 87B. The gene is intronless and appears to possess conventional 5' TATA, Cap and 3' polyadenylation signals. A single transcript, approximately 1kb in size, appears to be expressed at high levels in all developmental stages examined. When this gene is overexpressed using various upstream GAL4 driver systems, no striking phenotypes are observed; however, we detect bristle morphology defects in some progeny. The gst-3 gene does not appear to be essential, based upon our observation that mutant flies homozygous for an EP element insertion 5' to the TATA box produce little or no detectable gst-3 mRNA; these flies are viable and fertile at 25 and 29 degrees C. Nevertheless, the gst-3 gene appears to be evolutionarily conserved in other Drosophila species, suggesting that it may be functionally important.
我们在此报告从黑腹果蝇中克隆并鉴定出一个新的θ类谷胱甘肽-S-转移酶(GST)基因gst-3。其序列与先前鉴定的果蝇GST基因不同,Southern印迹显示基因组中没有其他密切相关的基因。原位杂交将该基因定位到2号染色体(55D),靠近gst-2(53F),并且与87B处的gst-D簇明显分开。该基因无内含子,似乎具有常规的5' TATA、帽和3' 聚腺苷酸化信号。一个大小约为1kb的单一转录本似乎在所有检测的发育阶段都高水平表达。当使用各种上游GAL4驱动系统过表达该基因时,未观察到明显的表型;然而,我们在一些后代中检测到刚毛形态缺陷。基于我们的观察,即TATA框上游5' 插入EP元件的纯合突变果蝇产生很少或没有可检测到的gst-3 mRNA,gst-3基因似乎不是必需的;这些果蝇在25和29摄氏度下可存活且可育。尽管如此,gst-3基因在其他果蝇物种中似乎在进化上是保守的,这表明它可能在功能上很重要。