title: Perturbation of local endogenous expression by insertion of Pol I expression constructs into the genome of Trypanosoma brucei.
abstract: Manipulation of gene expression is a common tool for the elucidation of biological function. Here we investigated the effects of over-expression in trypanosomes of a small GTPase, TbRABX1, using 2D gel electrophoresis and mass-spectrometry. An over-expression construct was targeting to the tubulin locus of chromosome I for stable integration and expression. Unexpectedly we observed alterations to the expression of gene products, i.e., tubulin, from surrounding regions of the genome; this effect was shown to be general and not dependent on the identity of the ectopic gene being expressed. These data suggest that local perturbation of the genome by insertion of DNA constructs can have wider impacts on gene expression, which need to be monitored.
authors.affiliation: Department of Biological Sciences, Imperial College, London SW7 2AY, UK.
authors.forename: Vivek
authors.initials: V
authors.lastname: Dhir
authors.forename: Clare L
authors.initials: CL
authors.lastname: Allen
authors.forename: Mark C
authors.initials: MC
authors.lastname: Field
mesh_terms: D000818:Animals; D015153:Blotting, Western; D004256:DNA Polymerase I / Q000235:genetics* / Q000502:physiology; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000235:genetics*; D018503:Genome, Protozoan; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000502:physiology*; D014404:Tubulin / Q000235:genetics / Q000502:physiology; D020691:rab GTP-Binding Proteins / Q000235:genetics* / Q000502:physiology
chemical_list: D014404:Tubulin; D004256:DNA Polymerase I; D020691:rab GTP-Binding Proteins