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dc.contributor.authorPrieto, Ignacio-
dc.contributor.authorMéndez Cormán, Enrique-
dc.contributor.authorSalas, Margarita-
dc.identifier.citationGene 77(2): 195-204 (1989)es_ES
dc.description.abstractUnit-length φ29 DNA was not synthesized after restrictive infection of Bacillus subtilis with the φ29 mutant sus 1(629) indicating that the phage φ29 protein p1 is needed for the viral DNA replication. Sequencing of the ORF-6 of mutant sus1 (629) showed that a C in the wild-type (wt) phage had been changed to a T at nt position 19 of the ORF-6, giving rise to a TAA ochre codon, indicating that this ORF corresponds to gene 1. ORF-6 was cloned in plasmid pPLc28 under the control of the pL promoter of phage λ and, after induction, a protein of about 10 kDa was overproduced, which was absent in the corresponding cells harbouring a recombinant plasmid with the sus1 (629) mutation, indicating that the 10-kDa protein is the product of gene 1. In addition, a protein of lower Mr was synthesized after induction of the cells harbouring recombinant plasmids with the wt or the sus1 (629) DNA. Both proteins were purified and characterized by N-terminal sequence determination and amino acid analysis. The low-Mr protein, named Δ1, has a size of 6 kDa and corresponds to an internal in-phase initiation event in ORF-6.es_ES
dc.description.sponsorshipThis in- vestigation has been aided by research grant 5 ROl GM27242-08 from the National Institutes of Health, by grant No. 3325 from Comision Asesora para la Investigation Cienthica y TCcnica and by a grant from Fondo de Investigaciones Sanitarias. I.P. was a recipient of a postdoctoral fellowship from the Spanish Research Council.es_ES
dc.subjectRecombinant DNAes_ES
dc.subjectPhage λ pL promoteres_ES
dc.subjectProtein-primed DNA replicationes_ES
dc.titleCharacterization, overproduction and purification of the product of gene 1 of Bacillus subtilis phage ø29es_ES
dc.description.peerreviewedPeer reviewedes_ES
dc.contributor.funderNational Institutes of Health (US)-
dc.contributor.funderComisión Asesora de Investigación Científica y Técnica, CAICYT (España)-
dc.contributor.funderInstituto de Salud Carlos III-
dc.contributor.funderConsejo Superior de Investigaciones Científicas (España)-
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