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We have synthesized wild type DNA binding protein (DBP) of adenovirus type 5 (Ad5) and several truncated forms of this protein by a combination of in vitro transcription and translation. The proteins obtained were tested for binding to a single-stranded DNA-cellulose column. It could be shown that for binding of in vitro synthesized wild type DBP, it is necessary to add zinc ions to the in vitro translation system. Binding studies with the truncated proteins revealed that deletion of the carboxyl-terminal 46 amino acids abolishes DNA binding.