Soysa Radika, Tran Khoa D, Ullman Buddy, Yates Phillip A
Department of Biochemistry and Molecular Biology, Oregon Health & Science University, Portland, OR 97239, USA.
Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR 97239, USA.
Mol Biochem Parasitol. 2015 Dec;204(2):89-92. doi: 10.1016/j.molbiopara.2016.01.008. Epub 2016 Feb 2.
We have designed a novel series of integrating ribosomal RNA promoter vectors with five incrementally different constitutive expression profiles, covering a 250-fold range. Differential expression was achieved by placing different combinations of synthetic or leishmanial DNA sequences upstream and downstream of the transgene coding sequence in order to modulate pre-mRNA processing efficiency and mRNA stability, respectively. All of the vectors have extensive multiple cloning sites, and versions are available for producing N- or C- terminal GFP fusions at each of the possible relative expression levels. In addition, the modular configuration of the vectors allows drug resistance cassettes and other components to be readily exchanged. In toto, these vectors should be useful additions to the toolkit available for molecular and genetic studies of Leishmania donovani.
我们设计了一系列新型的整合核糖体RNA启动子载体，具有五种逐渐不同的组成型表达谱，覆盖范围达250倍。通过在转基因编码序列的上游和下游放置合成或利什曼原虫DNA序列的不同组合来实现差异表达，以便分别调节前体mRNA加工效率和mRNA稳定性。所有载体都有广泛的多克隆位点，并且有不同版本可用于在每个可能的相对表达水平产生N端或C端GFP融合蛋白。此外，载体的模块化配置允许耐药盒和其他组件易于交换。总体而言，这些载体应该是对用于杜氏利什曼原虫分子和遗传研究的工具集的有用补充。
