Ljubljana/results

Results

Before testing the full functional devices we had to do some research on individual BioBricks to prove their expression and activity of their protein products in mammalian cell cultures. Preliminary testing included control of surface expression of transmembrane receptors with flow cytometry and testing and optimization of the T7 promoter system in mammalian cells.
We continued with testing of individual subsystems; fluorescent reporter proteins were used to prove the functionality of subsystems with confocal microscopy and luminescence for luciferase reporters. Western blotting was used to proteolytic cleavage of test split and TEV protease subsystems. Final stage was testing of effector protein activity – caspase 3. Its expression was determined by ELISA test and its activity, resulting in cell apoptosis was detected by flow cytometry.
We have shown that anti-HIV defense based on HIV entry or its activity in infected cell can lead to transcription and activation of effector proteins that either trigger apoptosis or enhance antiviral immune response.
Detailed information on results of our experiments can be found in the following chapters.