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Updated: Aug 8, 2026
Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
Cloning and functional analysis of cDNA encoding the hamster Bcl-2 protein
M T Tomicic1, M Christmann, B Kaina
1Division of Applied Toxicology, Medical Faculty, Institute of Toxicology, University of Mainz, Obere Zahlbacher Strasse 67, Mainz, D-55131, Germany.
Abstract:
We have cloned cDNA encoding hamster Bcl-2 protein from total RNA of CHO-9 cells by RT-PCR using oligonucleotide primers sharing homology with the sequence of mouse and rat bcl-2. The fragments spanning the total coding region were cloned into pCR4-TOPO and sequenced for verification. The hamster bcl-2 cDNA has a size of 711 nucleotides and encodes a polypeptide of 236 amino acids. Hamster Bcl-2 shares 95.8 and 88.6% similarity with mouse and human Bcl-2, respectively. Northern blot analysis revealed a single 7.5 kb bcl-2 transcript in hamster (CHO-9), mouse (BK4), and rat (H5) cells and a 8.5 kb bcl-2 mRNA in human (HeLa MR) cells. The bcl-2 cDNA (771 bp) was recloned into pcDNA3 and the recombinant construct was transiently transfected into MGMT-deficient HeLa MR cells. Expression of hamster Bcl-2 rendered the cells more resistant to MNNG-induced cytotoxicity, which is consistent with the anti-apoptotic function of Bcl-2.
