Nucleotide sequence analysis and expression of the minimum REPI replication region and incompatibility determinants of pColV-K30.

Abstract:

:We sequenced the minimum REPI replication region and the incompatibility determinants of pColV-K30. The minimum replication region contains an open reading frame which corresponds to a 35-kilodalton (kDa) protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis with maxicells transformed with a number of deletion derivatives demonstrated that this replication region encodes a 39-kDa protein and also established the direction of transcription of the RepI protein gene. The 39-kDa polypeptide was identified as the trans-acting factor essential for replication of REPI-containing plasmids. A translated region of the nucleotide sequence of the RepI protein gene showed homology with the helix-turn-helix binding domains of a number of DNA-binding proteins and also with other plasmid replication proteins. Further nucleotide analysis of the REPI region revealed the presence of direct and inverted repeat sequences in the incE, incF, and ori regions. The REPI ori also contained a perfect DnaA-binding site in addition to a high frequency of occurrence of the DNA adenine methylation (dam) site 5'GATC3'.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Perez-Casal JF,Gammie AE,Crosa JH

doi

10.1128/jb.171.4.2195-2201.1989

subject

Has Abstract

pub_date

1989-04-01 00:00:00

pages

2195-201

issue

4

eissn

0021-9193

issn

1098-5530

journal_volume

171

pub_type

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