کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5916053 1163355 2008 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Single-locus targeting constructs for reliable regulated RNAi and transgene expression in Trypanosoma brucei
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شناسی مولکولی
پیش نمایش صفحه اول مقاله
Single-locus targeting constructs for reliable regulated RNAi and transgene expression in Trypanosoma brucei
چکیده انگلیسی
A major obstacle to reproducible expression of recombinant transcripts lies in the epigenetic effects of the flanking chromatin following integration. We previously presented a strategy to overcome this problem in bloodstream form Trypanosoma brucei, using a reporter to identify a ribosomal-spacer locus that supports optimal expression and then marking that locus for subsequent targeting. Advantages include elimination of variable-expression position-effects and the easy confirmation of correct integration. We now report a set of validated constructs that exploit this system for expression of dsRNA or recombinant protein. The current construct-set allows expression of intramolecular dsRNA for RNA interference knockdown or expression of proteins that can incorporate c-Myc epitope(s) or a fluorescent-tag for subcellular localisation, interaction and/or other functional analysis. The constructs are integrated at a single, marked locus and deliver reliable and reproducible expression.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Molecular and Biochemical Parasitology - Volume 161, Issue 1, September 2008, Pages 76-79
نویسندگان
, ,