Investigation of the effects of deletion of transcription factor binding sites in human BAX gene 5′ noncoding sequence on gene expression

In this study, the human BAX gene, a proapoptotic gene, was identified the 2000 bp 5′ noncoding sequence by using the Ensembl database. According to the functional features of this region, 1400 bc, 930 bc and 390 bc fragments were separated by PCR method. The amplified fragments were transferred to the pGEM-T Easy vector and transferred to Escherichia coli JM109 bacteria. Subsequently, the obtained regions were transferred to the luciferase vector (pGL-3 basic vector). Purified plasmids were transfected into MCF-7 cells. Oluşturulan konstraktların lusiferaz aktivitesi, kontrol vektörüne göre ölçümü yapıldığında 930 bç'lik parçanın en yüksek aktivite gösterdiği tespit edildi. Using the Genomatix database, 930 bp fragments were divided into 797 bp, 673 bp, 230 bp and 185 bp fragments according to the location of the SWI / SNF transcription factor binding sites. Çoğaltılan parçaların lusiferaz aktivitesi kontrol vektörü ile kıyaslandığında en yüksek aktivitenin 673 bç'lik bölgede bulunan SWI-SNF3 ve SWI-SNF4 olarak adlandırdığımız transkripsiyon faktör bağlanma bölgelerinde olduğu tespit edildi. Thus, site-specific mutagenesis on transcription factor binding sites detected in later studies will allow interaction with different transcription factors on the 5'non-coding sequence.