Characterization of the E.coli K12 strain AB1157 as impaired in guanine/xanthine metabolism

The widely used E. coli K12 strain AB1157 is impaired in guanine (xanthine) metabolism. Mutants blocked in purine biosynthesis before the stage of inosine monophosphate synthesis do not grow on external guanine or xanthine.The genetic nature of the Gua/Xan lesion is a deletion in the chromosome that covers the pro A gene. The lesion causes reduced uptake of guanine.     

Some group N plasmids makeEscherichia coli K-12 strain AB1157 dependent on exogenous purine

Escherichia coli K-12 strain AB1157 given the conjugative group N plasmid R46 (or its derivative, pKM101, or plasmid R384N) grew only very slowly on defined medium containing the known growth-factor requirements of AB1157, which do not include any purine. Addition of adenine or hypoxanthine (or their nucleosides) restored normal growth; guanine and xanthine (and their nucleosides) were ineffective, because of thegpt defect caused by deletionproA2. Variants of AB1157(R46) able to grow rapidly on defined medium without purine were tetracycline-sensitive and/or transfer-defective; an, R46 gene,slo, causing purine auxotrophy, is inferred to be betweentet andtra.     

Phenotypic variations in strain AB1157 cultivars of Escherichia coli from different sources.

The purpose of this note is to alert users of Escherichia coli AB1157 and its derivatives to a potentially significant difference in cultivars from various sources. The difference we find is in the ability to host an infection by coliphage 186 after UV irradiation of the host cell.     

On the protection of E. coli against radiation lethality by ascorbate combined with tetracycline.

The radioprotective action in E. coli ATCC 9637 of ascorbate added to media containing the weak sensitizer, tetracycline (effect described by Pittillo and Lucas (1967)), was found to be dependent on the presence of metal catalysts of the autoxidation of ascorbate. Thus, the protective action of ascorbate + tetracycline as well as the rate of autoxidation of ascorbate in this mixture were enhanced by 0.1 micron Cu, and these effects were counteracted by pyrophosphate probably through chelation of iron that contaminates phosphate. A suppression of metabolism is apparently involved in the combined action as judged by the decrease of incorporation of labelled uridine.     

Analysis of ultraviolet light-induced suppressor mutations in the strain of Escherichia coli K-12 AB1157: an implication for molecular mechanisms of UV mutagenesis

Summary Genetic analysis of histidine independent (His⁴) revertants induced by ultraviolet light in the his-4 E. coli strain AB1157 was carried out: 83% carried ochre (UAA) suppressor mutations and 17% carried back mutations to his ⁺ or (intragenic?) suppressors not detectably separable from his-4. Using the specialized transducing psu 2int ^– phage, which carries supE-supB, it was determined that 87% of the ochre suppressors mapped in the supE-supB region. We were able to deduce that 56% of these affected tRNA ₁ ^Gln by a CAATAA change in the tRNA gene while 31% affected tRNA ₂ ^Gln by TAGTAA change. Although we were unable to deduce the base substitution of the remaining 13%, the results indicated that most of the suppressor mutations are caused by a G:C to A:T transition.These results suggest that the high incidence of supE-supB region suppressor mutation in E. coli by UV would be a reflection of the general feature of UV mutagenesis; i.e. preferential induction of G:C to A:T transition in repairing nonparing DNA lesions.     

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.     

Secretion of a 107 K dalton polypeptide into the medium from a haemolytic E. coli K12 strain

Summary Certain E. coli K12 strains are able to secrete a plasmid encoded 107 K protein into the culture medium. During exponential growth of the cells this protein represents approximately 1% of total cell protein.The presence of the 107 K polypeptide was demonstrated through the fortuitous use of strain MC4100. This gave a largely protein-free culture supernatant, presumably due to minimal lysis of whole cells. Pulse-labelling experiments showed that the secretion of the 107 K polypeptide reached a maximum during the stationary phase of growth, where it represented substantially more than 1% of total cell protein. The 107 K polypeptide is coded by the haemolytic plasmid pHly167, and appears to be related to a previously reported intracellular precursor form of the -haemolysin (Goebel and Hedgpeth 1982). However, additional extracellular factors appear to be required for -haemolysin activity since several nonhaemolytic mutants still secrete this protein.     

Difference in frequency spectrum of extremely-low-frequency effects on the genome conformational state of AB1157 and EMG2 E. coli cells

The effect of weak extremely-low-frequency (ELF) magnetic fields (sinusoidal, 30 μT amplitude) on the genome conformational state (GCS) of E. coli mutant and wild type cells was studied by using the method of anomalous viscosity time dependency (AVTD) in the 6–37 Hz frequency range. We confirmed the existence of three resonance frequencies of 8.9, 15.5, and 29.4 Hz when mutant cells of K12 AB1157 strain were exposed. In the same frequency range, the wild type K12 EMG2 cells displayed only two effective windows, with resonance frequencies of 8.3 and 27 Hz. The resonance frequencies differed significantly (P < .001–.000001) in the strains studied, whereas other resonance parameters did not. It was concluded that mutations in the AB1157 strain resulted in a significant rearrangement in the ELF action spectrum, including the appearance of a new resonance. © 1996 Wiley-Liss, Inc.     

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.     

Isolation of streptothricin resistant mutants from E coli K12, strain A19

Mutants with various levels of resistance to streptothricin were isolated from Escherichia coli K12, strain A19 after mutagenesis with N-methyl-N-nitro-N-nitroso-guanidine and ethylmethane-sulfonate. Nourseothricin, a mixture of streptothricin F and D was the selection agent. Spontaneous resistant mutants could not be found. The streptothricin-resistant mutant E. coli A19 Stcr 2/2/1 shows cross-resistance to some of the aminoglycoside antibiotics investigated, but no cross-resistance to chloramphenicol and chlortetracyclin. These results indicate similar mechanisms of action of streptothricin and aminoglycoside antibiotics.     

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.     

Selective effect of p-aminobenzoic acid on mutagenesis: phenotypic analysis of the Arg+-revertants induced by N-nitroso-N-methylurea in the Escherichia coli K-12 AB1157 strain

According to the phenotypic analysis of Arg+ revertants in Escherichia coli K-12 AB1157, the specific mutational changes in bacterial cells under the action of MNU were registered. True and suppressor mutations of four phenotypic groups were noted. The quantity of mutants induced depended upon the DNA and protein syntheses in bacterial cells. The sublethal concentration of para-aminobenzoic acid markedly (10-50 fold) reduced the rate of mutagenic induction and changed the quantity relations of mutants within phenotypic groups.