Newfeld S J, Takaesu N T
Department of Biology, Arizona State University, Tempe, Arizona 85287-1501, USA.
Genetics. 1999 Jan;151(1):177-87. doi: 10.1093/genetics/151.1.177.
We have efficiently mobilized a phenotypically silent hobo transgene inserted within the cis-regulatory heldout region of the decapentaplegic (dpp) locus in Drosophila melanogaster. The goal of our experiment was to identify germline transmission of a local transposition event within the dpp locus that meets two specific criteria. First, excision of the hobo construct does not generate an adult mutant phenotype, suggesting minimal alteration to the original site of insertion. Second, we required a new insertion of the hobo transgene into the Haploinsufficient region of the locus approximately 25 kb away. Genetic and molecular criteria are used to evaluate candidate germlines. In a pilot study, this local transposition event occurred independently in two individuals. Both of the transposition events appear to be new insertions into the dpp transcription unit. One insertion is between the two protein-coding exons, and the other is in the 3'-untranslated region of exon three. Strains carrying these insertions are valuable new reagents for the analysis of dpp function and molecular evolution. These results further support the use of the hobo system as an important tool in Drosophila genetics.
我们已经有效地激活了插入在黑腹果蝇中decapentaplegic（dpp）基因座顺式调控保留区域内的一个表型沉默的hobo转基因。我们实验的目的是鉴定dpp基因座内符合两个特定标准的局部转座事件的种系传递。第一，hobo构建体的切除不会产生成年突变体表型，这表明对原始插入位点的改变最小。第二，我们要求hobo转基因新插入到该基因座大约25 kb远处的单倍体不足区域。遗传和分子标准用于评估候选种系。在一项初步研究中，这种局部转座事件在两个个体中独立发生。这两个转座事件似乎都是dpp转录单元的新插入。一个插入在两个蛋白质编码外显子之间，另一个在外显子三的3'非翻译区。携带这些插入的菌株是用于分析dpp功能和分子进化的有价值的新试剂。这些结果进一步支持将hobo系统用作果蝇遗传学中的重要工具。
