Using genetic and metabolic engineering, we are able to modify metabolism of certain organisms. Filamentous fungi are one of the most biotechnologically important industrial microorganisms. The key characteristic that makes them so important, is their ability to produce numerous metabolites such as proteins, organic acids, enzymes and secondary metabolites. Trichoderma reesei has been regarded as a leading fungus for metabolic engineering, due to its ability to produce and to secrete high amounts of cellulases. Among the various molecular approaches, application of reporter gene is very important for studying and optimizing the heterologous protein production in the genome of the desired host. In order to establish an appropriate methodology for heterologous protein production in the genome of fungus T. reesei RUT-C30, we constructed and applied different vectors that contain reporter gene for green fluorescent protein (GFP). As a result, we obtained T. reesei transformants which can express reporter gene for GFP that was under the control of different promoters. We observed that the production of heterologous GFP protein was higher with the application of Avicel inductor and cbh1 promoter. Certain carbon sources strongly regulate the activity of the promoter of the cbh1 gene. With the application of spectrophotometric measurement of fluorescence, SDS PAGE, western blot and fluorescence microscopy, we successfully detected intracellular expression of GFP.
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