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Four mutations conferring recombination deficiency (recA 1, recA 13, rec-12, and rec-65) have been cotransduced with cysC and pheA; consequently they lie between 53 and 50 on the standard genetic map of Escherichia coli. The four mutations show different degrees of apparent dominance in transient rec+/reczygotes, and the degree of apparent dominance of a particular mutation was shown to be a characteristic of that mutation, not a reflection of other genetic differences in the original rec mutant strain. All four mutations map at similar distances from cysC and pheA and, despite the different degrees of apparent dominance, all may lie in the recA gene.

The IncP plasmid R68.45 and other plasmids carrying tandem repeats of the insertion sequence IS21 [= (IS21)2] produce replicon fusions via transposition at high frequencies in Escherichia coli and other gram-negative bacteria, whereas plasmids with a single IS21 copy, e.g. R68, give replicon fusions rarely. The 2131 bp nucleotide sequence of IS21 was determined; at the ends there were 11 bp inverted repeats with one mismatch. Two adjacent open reading frames, istA and istB, were located on one DNA strand of IS21. In E. coli maxicells, polypeptides of 46 kDa (the istA gene product) and 30 kDa (the istB gene product) were expressed by (IS21)2 plasmids, but not by IS21 plasmids. Genetic analysis of (IS21)2 plasmids indicates that the IS21-IS21 junctions form a promoter, which initiates transcription of the istAB operon in one of the two IS21 elements. A single IS21 element fused to an inducible external tac promoter expressed both proteins after induction, but did not promote effective replicon fusion, unless an IS21-IS21 junction (the preferred site for IS21 transposase action) was also present on the plasmid carrying the tac-IS21 construct. The sequences located between the IS21 elements in (IS21)2, 3 bp in R68.45 or 2 bp in pME28, were not recovered in the replicon fusion products. Homologous recombination between the directly oriented IS21 elements in the fusion products led to plasmids with a single IS21 insertion. Analysis of the latter showed that IS21 had a low, but not totally random specificity of insertion and created target duplications of 4 bp (occasionally 5 bp).(ABSTRACT TRUNCATED AT 250 WORDS)

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Eighty-four transfer-deficient mutants of Flac have been isolated; 27 of these bear amber mutations and I mutant is temperature-sensitive. All the mutants transfer between 10-2 and < 1O-5% as well as wild-type Flac, all are curable by acridine orange treatment, and all are resistant to the female-specific phage MI,. Some of the mutants are partially sensitive to female-specific phage tau. Sixtythree of the mutants are resistant to the male-specific phages f1, f2, and Q,B; 15 are resistant only to f2; and 6 are sensitive to all three male-specific phages. Most of the mutants are still poor recipients in conjugation, but four of the mutants resistant to fl, f2, and Q# have become good recipients. Those mutants resistant to all three male-specific phages do not seem to make F-pili.

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The conjugation regions of IncF plasmids are closely related in that they share extensive DNA homology, and that they specify related pili. Variations between individual conjugation gene products of different IncF plasmids have, however, been noted. We have extended these observations by carrying out a systematic survey of twelve such plasmids, to examine the numbers and the groupings of the plasmid-specific alleles of several genes required for conjugation and its control.Using vector plasmids carrying cloned origins of transfer (oriT), four different specificities were recognized, and these were correlated with the specificities of the genes with products that may act at this site (traM, traY and traZ). The traY gene is the first gene of the major transfer operon, and is therefore located close to the site at which the traJ protein acts to induce expression of the operon: correspondingly, correlation was observed between the oriT/traMYZ and traJ specificities in most of the plasmids. In turn, traJ is negatively regulated by the finO and finP products acting in concert: the finO product was relatively non-specific, but six finP alleles were identified, again with , specificities correlated with those of traJ. Our explanation for this unexpectedly large number of finP alleles derives from the concept that the finP product is an RNA molecule rather than a protein. Although the conjugative pili encoded by IncF plasmids are closely related, they confer different efficiencies of plating of the various F-specific bacteriophages. We distinguished four groups on this basis, presumably resulting from differences in the primary amino-acid sequences of the pilin proteins. These groups could be related to the surface exclusion system specificities, consistent with the hypothesis that surface exclusion acts at least in part by preventing interaction between the pilus and the recipient cell surface.From these data, information about the evolutionary relationships between the twelve IncF plasmids can be deduced.