Role of Bovine Bcl2A1 Gene in Staurosporine-Induced Apoptotic Cell Lines

B-cell lymphoma 2 - related protein A1 (Bcl2A1) is a member of a gene set that plays a critical role in regulating apoptosis of mammalian cells. The family is divided into 2 groups: 1. anti-apoptotic genes Bcl-2, Bcl-xL, Bcl2A1 and 2. pro-apoptotic genes Bid, Bax, Bak. In this study, the ORF encoding the 175 amino acid bovine Bcl2A1 protein was amplified by reverse transcription polymerase chain reaction , using specific primers deduced from the bovine Bcl2A1 mRNA sequence and which also contained appropriate restriction endonuclease cleavage sites for cloning. The product was then cloned into plasmid pcDNA3+ to construct the eucaryotic expression plasmid pcDNA3.Bcl2A1. Hela, L11, Vero and WSL cell lines were used to investigate apoptosis induced by staurosporine. Monitoring of cellular DNA fragmentation revealed that incubation with staurosporine at a concentration of 2 mu M results in a sufficient level of apoptosis induction in Hela, L11, WSL cell cultures after 6 hours and in Vero cell cultures after 12 hours.