Cloning and expression in Escherichia coli of the gene coding for the protein linked to the ends of Bacillus subtilis phage phi 29 DNA.

A phi 29 DNA fragment containing gene 3, coding for the 5'-terminal protein, and several other early genes has been cloned in a pBR322 derivative plasmid (pKC30) under the control of the pL promoter of bacteriophage lambda. Four polypeptides of Mr 27000, 18500, 17500 and 12500 were labelled with [35S]methionine after heat induction, accounting for about 15% of the de novo synthesized protein. The Mr 27000 and 12500 proteins were characterized as p3, the 5'-terminal protein, and p4, involved in the control of late transcription, respectively. Protein p3 synthesized in Escherichia coli was active in the in vitro formation of the initiation complex p3-dAMP when supplemented with extracts from Bacillus subtilis infected with a sus3 mutant.