Synthesis of paf-acether by E. coli K12

Paf-acether (platelet-activating factor) is one of the most potent mediator of inflammation released from and acting on most cells that participate in inflammatory diseases. Its molecular structure is 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine. Two metabolic steps are involved in its biosynthesis: the action of a phospholipase A2 on choline-containing membrane alkyl-ether lipids results in the production of lyso paf-acether and acetylation of the lyso compound by an acetyltransferase yields the biologically active molecule. Membrane alkyl-ether lipids can therefore be considered as potential precursors of paf-acether and their composition has been studied in various cell types. In this work, we investigated the presence of paf-acether in E. coli. Our results showed that paf-acether can be obtained from E. coli K12 under a variety of bacterial growth conditions. Paf-acether from E. coli exhibited the same physicochemical and biological characteristics as synthetic paf-acether and that from eucaryotic cells. Therefore, it appears that E. coli itself has the ability of producing paf-acether, a result that could be of some importance with respect to the pathogenesis of Enterobacteria and the use of E. coli in the recombinant DNA technology.     

Sequence of the malK gene in E.coli K12.

We present the sequence of gene malK which encodes a component of the system for maltose transport in E.coli K12. We also determined the position of deletion (S50) which fuses malK to the following gene lamB; the malK-lamB protein hybrid contains all of the malK protein. The mRNA corresponding to the last two thirds of gene malK could form stable stem and loop structures. The malK protein, as deduced from the gene sequence, would include 370 residues and correspond to a molecular weight of 40700. The sequence as well as sequence comparisons with the ndh protein of E.coli are discussed in terms of the location and function of the malK protein.     

Ultrastructure of LLR-mutants of E. coli K12]

The author studied the ultrastructure of two spherical E. coli K12 mutants (llr) obtained under the effect of N-nitroso-N-methylurea. Seven morphological types of cells differing from one another by shape, size and cytoarchitectonics were distinguished. Superficial structures of the majority of the cells were represented by the membranes of the cell wall and the cytoplasmic membrane of common structure. Some of the cells had only one membrane coat and a high electron optic density of the cytoplasm. Transitional forms of cells were also encountered. The ultrastructure of each morphological type in the population of the llr-mutants was described in detail. The capacity of the mutants to vacuolization, to the intra- and extracellular budding, and also the ability to form multiple membrane structures resembled analogous structures of stable L-forms of the Gram-negative microbes. The problems of morphological differentiation of the L-forms and of the llr-mutants, and also problems connected with the formation of the multiple membrane structures and small elemental bodies in the cells of the llr-mutants are discussed.     

IS4 at its chromosomal site in E. coli K12

Summary IS4 DNA has been isolated in pure form. Hybridization of this DNA against restricted DNA of several E.coli K12 strains by Southern's blotting technique has shown that, in most strains, only one copy of IS4 is present. Though the restriction fragments around this site differ in size, IS4 can be shown always to be located at the same site. In one strain, one additional copy has been found in a new location. In this strain, IS4 in its original location has been retained.     

Lethal effects of high-voltage pulses on E. coli K12

Summary The lethal effects of high-voltage capacitor-discharges in suspensions of E. coli K12 with varying electrolytes have been examined. A reduction of more than 99.9% of living cells, dependent on the applied voltage could be proved. The bactericidal action is assumed to be due to direct effects of high electric fields. Electrolytically produced chlorine was shown to act as an additional toxic agent, when chloride is present in the treated medium. The relative survival rate of bacteria has been found to depend also on the concentration of cells during pulse treatment.     

Dominance study with rifampicin-resistant mutants of E. coli K 12

Summary The location of the characters for rifampicin-resistance in E. coli linked to the arg E marker was confirmed by mating experiments with rifampicin-resistant Hfr C mutants and a rifampicin-sensitive acceptor strain. During these tests, temporary dominance of rifampicin-sensitivity was observed. On the basis of the results obtained, the time required for complete phenotypic expression of rifampicin-resistance was determined, and the ratios of rifampicin-sensitivity to rifampicin-resistance in the merozygote phase of the cell were studied. It was found that the episomal marker of rifampicin-sensitivity influences the level of resistance in merodiploid cells, but does not fully restore the original sensitivity of the wild type.     

Alignment of Escherichia coli K12 DNA sequences to a genomic restriction map.

We use the extensive published information describing the genome of Escherichia coli and new restriction map alignment software to align DNA sequence, genetic, and physical maps. Restriction map alignment software is used which considers restriction maps as strings analogous to DNA or protein sequences except that two values, enzyme name and DNA base address, are associated with each position on the string. The resulting alignments reveal a nearly linear relationship between the physical and genetic maps of the E. coli chromosome. Physical map comparisons with the 1976, 1980, and 1983 genetic maps demonstrate a better fit with the more recent maps. The results of these alignments are genomic kilobase coordinates, orientation and rank of the alignment that best fits the genetic data. A statistical measure based on extreme value distribution is applied to the alignments. Additional computer analyses allow us to estimate the accuracy of the published E. coli genomic restriction map, simulate rearrangements of the bacterial chromosome, and search for repetitive DNA. The procedures we used are general enough to be applicable to other genome mapping projects.