Nucleotide sequence and gene organization of ColE1 DNA.

P T Chan,H Ohmori,J Tomizawa,J Lebowitz

doi:10.1016/s0021-9258(17)39438-3

Abstract

The primary structure of the plasmid ColE1 DNA has been determined. The plasmid DNA consists of 6646 base pairs (molecular mass of 4.43 MDa) and is 48.46% in GC content. The phi 80 trp insert of the composite plasmid of ColE1, pVH51, has also been determined. The determination of the nucleotide sequence of ColE1 DNA provides the basis for examining the relationships between the DNA sequence and the gene organization of the plasmid. The focus of this paper is to use this sequence data coupled with a review of the literature and our own work to examine the nine known functional regions of ColE1: imm (colicin E1 immunity), rep (replication function), inc (plasmid incompatibility and copy number control), bom (basis of mobility), rom (modulator of inhibition of primer formation by RNA I), mob (plasmid mobilization), cer (determinant for conversion of plasmid multimers to monomers), exc (plasmid entry exclusion), cea (structural gene for colicin E1), and kil (structural gene for the Kil protein).

Highlights

The primary structure of the plasmid ColE1 DNA has been determined
Considerable attention has been directed on the action of colicin El, the bactericidal protein responsible for the killing of other enteric bacteria.Interest in theseandother phenomena (chromosome mobilization, colicin E l immunity, plasmid entry exclusion, and theconversion of plasmid multimers to monomers to be described below [11,12,13,14,15,16]) makes the determination of the nucleotide sequence extremely valuable
Parts of the nucleotide sequence of ColEl DNA has been reported by different laboratories [17,18,19,20,21,22], we have determined over 90% of the nucleotide sequence of ColE1

Summary

11 Present address

Institute for Virus Research, Kyoto University, Kyoto 606, Japan. ColEl [23, 24] containing the DNA sequence found between residues 1 and 3371 and part of the 480 trp region. Since pVH51 and its derivatives have been used in DNA physical studies [25, 26], we determined the sequence of the 480 trp insert region (Fig. 2) so that the full sequence of this ColEl derivative would be available to investigators.

EXPERIMENTAL PROCEDURES

RESULTS

I 1 1 1 1 1I I II I II II I 1 1 1 I I i I II II II

I I1 I I I II I I I

DISCUSSION