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

Lee EH,Masai H,Allen GC Jr,Kornberg A

doi

10.1073/pnas.87.12.4620

subject

Has Abstract

pub_date

1990-06-01 00:00:00

pages

4620-4

issue

12

eissn

0027-8424

issn

1091-6490

journal_volume

87

pub_type

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