Cloning and functional analysis of cDNA encoding the hamster Bcl-2 protein.
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.
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Division of Applied Toxicology, Medical Faculty, Institute of Toxicology, University of Mainz, Obere Zahlbacher Strasse 67, Mainz, D-55131, Germany.
Christmann
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Kaina
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D000478:Alkylation; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002273:Carcinogens / Q000633:toxicity; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D008769:Methylnitronitrosoguanidine / Q000633:toxicity; D008969:Molecular Sequence Data; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D002273:Carcinogens; D018076:DNA, Complementary; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D008769:Methylnitronitrosoguanidine