Specific Primer for Human Bcl-2-Protein Expression Analysis

Abstract

Bcl-2 protein (B-cell lymphoma 2) is one of the proteins in the Bcl-2 protein family that are anti apoptotic and encoded by the BCL-2 gene. This protein controls the permeability of the outer membrane of the mitochondria, which plays a role in the release of cytochrome c and several other proteins that contribute to cell death. Bcl-2 protein plays a role in regulating cell death and regulating the balance between new and old cells. Several studies showed that changes in the structure and function of Bcl-2 play a role in cell apoptosis disorders, producing cancer cells. High expression of Bcl-2 protein is seen in lung, breast, lymph node and brain cancers. Analysis of Bcl-2 protein expression at the molecular level in cancer cell is needed to determine the expression level of this protein. This study aims to produce the Bcl-2 primer for qPCR method that specifically identifies the target gene, and can be used to see the BCL-2 gene expression level in cells. A BCL-2 sequence was obtained from the National Center for Biotechnology Information (NCBI). The next process is further analysis with Primer-BLAST (NCBI). Some primer candidates are produced and undergo another selection based on several criteria. Finally, one pair of potential primers was obtained for the reaction of qPCR to BCL-2. Primer specificity was tested with Basic Local Alignment Tools (BLAST) tools and result in specific primer for human BCL-2 gene. Annealing temperature optimization shows that the primer can react optimally at 59∘C.   Keywords: Bcl-2 protein, BCL-2 gene, qPCR, primer, gene expression, apoptosis