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Transfer gene expression during fertility inhibition of the Escherichia coli K12 sex factor F by the I-like plasmid R62.

Further understanding of how the FinQ fertility inhibition system of the I-like plasmid R62 inhibits transfer of the sex factor F has been gained by the use of a genetic assay for individual transfer gene products. The technique involved construction of a series of Flac plasmids carrying suppressible mutations in individual transfer genes together with a FinQ inhibitor-insensitive traQ mutation. The transfer of the Flac double mutants from a strain carrying wild-type Fhis and R62 then indicated the effect of R62-encoded transfer inhibition on the expression of individual F transfer genes. During such inhibition the products of genes traJ, traA, traE, traB and traC were present in quantities sufficient to permit efficient F transfer, whereas the levels of the traF, traH, traG and traD gene products were so reduced as to limit F transfer. These findings and a failure to obtain recombination between traC and traQ mutations suggest that the R62 fertility inhibition system terminates transcription of the transfer operon between traC and traF.

Coliphages↗

Analysis of mitochondrial ribosomal proteins of Saccharomyces cerevisiae by two dimensional polyacrylamide gel electrophoresis.

Proteins from mitochondrial ribosomes of Saccharomyces cerevisiae were analysed by a two dimensional gel electrophoresis method. Each ribosomal subunit revealed a reproducible characteristic pattern of protein components. The 37S small subunit contained 33 protein species with an average molecular weight of 27,300 daltons (ranging from 60,000 to 95000 daltons). The 50S large subunit showed 38 protein species with an average molecular weight of 23,000 (ranging from 41,000 to 10,000 daltons). Ribosomes from various sources were compared on the basis of protein composition.

DNA, Mitochondrial↗

Construction and restriction endonuclease mapping of hybrid plasmids containing Saccharomyces cerevisiae ribosomal DNA.

Fragments produced by partial digestion of Saccharomyces cerevisiae ribosomal DNA (rDNA) with the restriction endonuclease EcoRI were ligated in vitro to the bacterial plasmid RSF2124. The resulting hybrid plasmids were cloned in Escherichia coli. Three hybrid plasmids which contain at least one intact repetitive unit of the multiple, tandem sequences of the yeast rDNA genes have been further characterized. These plasmids have been used to construct a map of the EcoRI, SmaI, HindII and HindIII restriction sites in the individual repetitive units of yeast rDNA.

DNA↗

Suppression of polarity of insertion mutations within the gal operon of E. coli.

Phenotypic revertants of galOP::IS1 and galOP::IS2 mutations have been isolated after mutagenesis with nitrosoguanidine, they are probably caused by mutations in gene suA. The polarity suppressor mutations described in this study and a known mutation in gene suA isolated by D. Morse (Morse and Guertin, 1972) suppress polarity caused by IS1 more effectively than that caused by IS2 or IS4. Furthermore, suppressibility is influenced by the site and orientation of IS integration. The synthesis of the three enzymes in galOP::IS suA double mutants is constitutive and the ratio of the three enzymes is altered in comparison to the wild type. The reasons for constitutive synthesis of the galactose enzymes and for the altered ratio of enzyme synthesis are discussed.

DNA, Bacterial↗

Altered tetracycline resistance in pSC101 recombinant plasmids.

Investigation of tetracycline resistance genetically determined by the plasmid pSC101 and several recombinants of pSC101 containing EcoRI generated DNA fragments inserted at the EcoRI site has revealed significant differences in the phenotypic expression of that resistance. The altered phenotypes of the recombinant plasmids may be the result of the location of the EcoRI site of pSC101, which has been determined to be near the genetic elements involved with tetracycline resistance.

Dose-Response Relationship, Drug↗

Respiratory repression and the stability of the mitochondrial genome.

The variation in sensitivity of the mitochondrial genome of Saccharomyces cerevisiae to ethidium bromide-induced petite mutation in response to changes in glucose concentration has been studied. Growth in high glucose considerably depressed the mutation rate, whilst small variations are observed in response to step-up or step-down in glucose concentration. Variations in mitochondrial DNA and respiratory activity during the mutagenic process are described. Effects of non-metabolizable sugars which repress mitochondrial biogenesis and a number of antimitochondrial drugs have been investigated. The results are discussed in terms of possible mechanisms of modulation of the mutation rates.

DNA, Mitochondrial↗

Replication of the mini-R1 plasmid Rsc11 and Rsc11 hybrid plasmids.

Replication of the multicopy mini-R1 plasmid, Rsc11, is dependent on host replication functions dna A, B, C, E and G but independent of polA1. Chloramphenicol immediately stops its replication. A stable relaxation complex is not formed. Composite plasmids were constructed with Rsc11 and other small replicons like pSC101, ColE1 and mini-ColE1. In all combinations the amount of hybrid plasmid DNA in the cell never exceeds the amount of Rsc11 DNA itself. This leads to varying copy numbers of the hybrid plasmids depending on the size of the second plasmid. Replication of the composite plasmids proceeds probably always under the control of the Rsc11 part although the second replicon is still functional. The composite plasmids are incompatible with both the parent replicons.

Chloramphenicol↗

Genetic analysis of unequal transmission of the mitochondrial markers in Saccharomyces cerevisiae.

The presence of mitochondrial sex factor, omega, was demonstrated in haploid strains of yeast Saccharomyces cerevisiae which came from our laboratory. Transmission and recombination of the mitochondrial genes (CR/CS, ER/ES and OR/OS), conferring the resistance/sensitivity to chloramphenicol, erythromycin and oligomycin, respectively, were non-polar in homosexual crosses and highly polar in heterosexual crosses. Different results were obtained in crosses involving an erythromycin resistant mutant G706E11 (CSEROS) which was found to contain cellular DNA of diploid level. This strain was omega- and showed no alleles from G706E11 (CS, ER and OS) were transmitted to the zygote progeny in preference to the CR, ES and OR alleles. When crossed to omega+ haploid strains, there was a highly polar recombination, but no transmission was seen for the E and O alleles. Polar transmission of markers from omega+ haploid parental strain, characteristic of heterosexual crosses, was noticed only for the C allele. The crosses of G706E11 to omega+ haploids featured an increase in the recombination frequency. The values of % suppressiveness of sigma- petite mutants were relatively low when determined by crossing to G706E11 or to sigma+ diploid strain M2-8C rather than by crossing to sigma+ haploid strains, indicating that there is a positive correlation between the polar transmission of drug resistance markers and the suppressiveness degrees. Genetic mechanism of the anomalous behaviors if mitochondrial genes in crosses involving G706E11 was discussed and interpreted as due to an unbalanced supply of mitochondrial genomes from parental strains.

Alleles↗

Integrative suppression of a dnaA mutation in Salmonella typhimurium.

Integrative suppression of a dnaA mutation in Salmonella typhimurium may result from the integration of F'lac or F'his into the chromosome in the left hand side of the chromosomal map. The suppressed revertants resulting from this integration do not contain DNA of the F' elements in the covalently closed circular (CCC)1 form but still contain the CCC DNA of the cryptic LT2 plasmid. Two suppressed revertants isolated from dnaA/F- strains were found in which the suppression of dnaA character was accompanied by the loss of CCC DNA from the cell lysates. From one of these revertants a segregant was isolated in which the return to the dnaA phenotype was accompanied by the reappearance of CCC DNA in the cell lysate. It is suggested that the cryptic plasmid may integrate into the chromosome of S. typhimurium and this integration may result in suppression of the dnaA mutation. Additional evidence suggesting that the cryptic plasmid controls its own initiation of replication independently of the function of the chromosomal dnaA gene is supplied by the results of the determination of incorporation of labelled thymidine into CCC DNA of the dnaA1 strain at the nonpermissive temperature.

Acridines↗

Mobilization of plasmid-borne drug resistance determinants for transfer from Pseudomonas aeruginosa to Escherichia coli.

RSU2, a plasmid transmissible between strains of P. aeruginosa but not to Escherichia coli can be mobilized by R751. Conjugatants receive a single plasmid composed of DNA from both R751 and RSU2 which has the compatibility properties of a member of group P (like R751). Study of this fusion plasmid suggests that the failure of RSU2 to transfer into enterobacteria is due to an inability to replicate in these bacteria. The fusion plasmid replicates using the genes of R751.

Ampicillin↗