Influence of the reporter vector backbone on 2-deoxyglucose dependent promoter activation

Aleksandra Solecka; Aleksandra Wielento; Joanna Jurczak; Weronika Sowińska; Mateusz Wawro; Aneta Kasza

doi:10.18388/abp.2020_5356

Aleksandra Solecka Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland https://orcid.org/0000-0003-4580-9850
Aleksandra Wielento Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland https://orcid.org/0000-0002-3657-3164
Joanna Jurczak Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland https://orcid.org/0000-0002-2424-1162
Weronika Sowińska Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland https://orcid.org/0000-0002-9549-9347
Mateusz Wawro Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland https://orcid.org/0000-0002-5964-3639
Aneta Kasza Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland https://orcid.org/0000-0001-9813-1567

DOI: https://doi.org/10.18388/abp.2020_5356

Abstract

Reporter vectors are very often used to investigate mechanisms responsible for regulation of promoter activity. Since their first generation, many new variants were constructed to increase sensitivity and reduce background signal. However, these tools are still imperfect and can generate false results. We have found that depending on the backbone of the reporter vector, pGL3 or pGL2, different results are obtained for a eukaryotic promoter’s activation by metabolic changes. These observations were done in the course of investigation of the MMP2 (matrix metalloproteinase-2) promoter regulation in response to inhibition of glycolysis.