Functional analysis of human bax gen promoter region

Cancers often result in a number of genetic changes in human life. Oncogenes and tumor suppressor genes are genes that control growth. Oncogenes encode proteins that normally stimulate growth. Malignant transformation occurs as a result of over-synthesis, translocations and mutations. Tumor suppressor genes normally encode proteins that inhibit growth. Undesirable conditions can occur if deletions occur in these genes, if the transcription event does not occur and if protein inactivation results in loss of function. In this study, the characterization of the proapoptotic Bax gene and the functional analysis of the promoter region, which have an important potential in the apoptosis-free mechanism, were performed. For this purpose, the 5000 bp promoter region of Bax gene was identified primarily by using bioinformatics programs and various databases. However, the overall analysis of our study was performed on a 2000 bp promoter fragment that we thought was functional. In this context, first, a genome transcription initiation site, a TATA binding box and probable p53 binding sites were identified. Later literature reviews and software programs were used to identify possible transcription attachment regions and motifs on this region. As a result of the analysis studies made, these 2000 bp parts were divided into four sub-parts according to their functional properties. And each fragment was cloned into the pGEM-T Easy vector. In this context, human genomic DNA was first purchased. PCR was performed with primers designed specifically for each region, and each region was obtained in pure form. The pure bands of each region were gel-extracted and transformed into the Escherichia coli JM109 bacterium by cloning into the pGEM-T Easy vector. Plasmid isolation was performed from each cloned fragment.