PMID:2162050
| Citation |
Lee, EH, Masai, H, Allen, GC Jr and Kornberg, A (1990) The priA gene encoding the primosomal replicative n' protein of Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 87:4620-4 |
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| 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. |
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| Keywords |
Amino Acid Sequence; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Base Sequence; DNA Replication; Escherichia coli/genetics; Genes, Bacterial; Genetic Vectors; Genomic Library; Molecular Sequence Data; Oligonucleotide Probes; Plasmids; Sequence Homology, Nucleic Acid |
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