title: The priA gene encoding the primosomal replicative n' protein of Escherichia coli.
abstract: The Escherichia coli gene encoding protein n' has been isolated and named priA for primosomal protein A. Protein n' is absolutely required for the conversion of single-stranded phi X174 DNA to the duplex replicative form in an in vitro-reconstituted system. The gene maps to 88.7 minutes on the chromosome adjacent to the cytR locus. Soluble protein extracts from cells harboring the priA gene on a multicopy plasmid contained 45-fold more n' replication activity than wild-type extracts. Enhanced overproduction of greater than 1000-fold was achieved by replacing the natural Shine-Dalgarno sequence with that of the phage T7 phi 10 gene and placing this priA under the control of the T7 phage promoter and RNA polymerase. The priA sequence reveals a 732-amino acid open reading frame and a nucleotide-binding consensus site consistent with the size and ATPase activity of the purified protein. The gene for protein n has been named priB and the putative gene for protein n", priC.
authors.affiliation: Department of Biochemistry, Beckman Center, Stanford University, CA 94305-5307.
authors.forename: E H
authors.initials: EH
authors.lastname: Lee
authors.forename: H
authors.initials: H
authors.lastname: Masai
authors.forename: G C
authors.initials: GC
authors.lastname: Allen
authors.forename: A
authors.initials: A
authors.lastname: Kornberg
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D015698:Genomic Library; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid
chemical_list: D001426:Bacterial Proteins; D015345:Oligonucleotide Probes