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The mRNA start site of a cloned rainbow trout protamine gene (TPG-3) has been localised using S1-nuclease mapping and primer extension of in vivo synthesised trout testis poly A+-RNA. The presumptive cap site occurs within an AT-rich region, only 14 nucleotides from the start of the protein-coding sequence. Transcription of this protamine gene in vitro, using the Hela whole-cell extract system, generates products initiated at the same nucleotide as that used in vivo. In vitro transcription is abolished by deletion of sequences between -20 and -48, within which is a canonical TATA-box having an llbp homology with the strong chick conalbumin and Adenovirus-2 major late promoters (CTATAAAAGGG).

The localisation of the 5′-termini ofin vivoandin vitrotranscripts of a cloned rainbow trout protamine gene