The influence of radiomimetic substances on deoxyribonucleic acid synthesis and function studied in escherichia coli/phage systems - IV. Colony-forming ability and escherichia coli strain B and two of its mutants following alkylation

1959 ◽  
Vol 151 (942) ◽  
pp. 129-147 ◽  
Keyword(s):  
Escherichia Coli ◽  
Chemical Reactivity ◽  
Alkylating Agents ◽  
Chemical Change ◽  
Acid Synthesis ◽  
Coli Strain ◽  
Ribonucleic Acids ◽  
Significant Difference ◽  
Synthetic Capacity ◽  
And Function

Escherichia coli strain B , its mutant B/r and a new mutant, designated B/HN 2, have been employed in a study of the effect of alkylating agents upon the survival of colony-forming ability and phage-synthetic capacity. This has been done against the background of our earlier work upon phage and that of other workers upon bacteria, employing both alkylating agents and radiations. The sensitivity of B toward all the compounds now studied was greater than that of the other two strains as regards colony-forming ability, whereas all three strains showed a similar sensitivity in regard to capacity. Survival curves of all strains treated with monofunctional agents were of a so-called ‘multi-hit’ type, whereas those for bifunctional compounds were downwardly concave. The response to di(2-ehloroethyl) methylamine ( HN 2) was complicated by the chemical change undergone by this substance in aqueous solution, as was shown by a comparison of fresh and aged solutions and of the effect of different cultural conditions prior to treatment. As with radiations, phage-synthetic capacity was considerably less sensitive to alkylation than colony-forming ability, whilst this sensitivity was essentially the same for the three strains. No significant difference was found between the effect of HN 2 on the capacity of B for T 2 and for T 7. The capacity of B/r for T 2 was more sensitive to treatment by a bifunctional agent than by a monofunctional agent of similar chemical reactivity. It is suggested that this may implicate ribonucleic acids as the reactive substrate essential to capacity.

The influence of radiomimetic substances on deoxyribonucleic acid synthesis and function studied in Escherichia coli / phage systems - III. Mutation of T 2 bacteriophage as a consequence of alkylation in vitro : the uniqueness of ethylation

1959 ◽  
Vol 150 (941) ◽  
pp. 497-508 ◽  
Keyword(s):  
Escherichia Coli ◽  
Burst Size ◽  
Alkylating Agents ◽  
Direct Consequence ◽  
Acid Synthesis ◽  
Deoxyribonucleic Acid Synthesis ◽  
Methane Sulphonate ◽  
Average Burst Size ◽  
And Function

Of a comprehensive set of alkylating agents tested, only two, namely, ethyl methane sulphonate and diethyl sulphate, have been found so to interact with T 2 bacteriophage that cells of Escherichia coli , infected with phage treated extracellularly, manifest a considerably increased likelihood of yielding mutated phage. Since this increase can occur where the infective titre of the phage and the latent period and average burst size of the infected bacteria remain unchanged, it is considered that the increased mutation rate is a direct consequence of the chemical treatment, although the alkylation itself does not constitute the mutation. A study of the manner of inactivation of the phage by these agents has not revealed any characteristic difference between ethylation and other alkylations which could be held to account for its apparent uniqueness.

The influence of radiomimetic substances on deoxyribonucleic acid synthesis and function studied in Escherichia coli / phage systems V. A comparison of the inactivation of phages T 2, T 7 and two strains of T 4 by alkylating agents

1959 ◽  
Vol 151 (942) ◽  
pp. 148-155 ◽  
Keyword(s):  
Deoxyribonucleic Acid ◽  
Osmotic Shock ◽  
Alkylating Agents ◽  
Acid Synthesis ◽  
Acid Moiety ◽  
Rate Of Penetration ◽  
Injection Process ◽  
Deoxyribonucleic Acid Synthesis ◽  
And Function ◽  
Escherichia Coli Phage

The sensitivity of phage T 7 to epoxides and freshly prepared solutions of di(2-chloroethyl) methylamine ( HN 2) was identical with that of T 2. T 7, however, proved considerably the more sensitive to ethylenimine and to aged solutions of HN 2. It was considered that this was due to the cationic nature of these latter agents affecting the rate of penetration into the phage heads, and that the susceptibility of T 2 and resistance of T 7 to osmotic shock was a parallel phenomenon. Confirmation was afforded by the fact that a strain of T 4 sensitive to osmotic shock behaved like T 2, and a resistant strain of T 4 like T 7. These results, together with others previously reported, are believed to offer very strong evidence that inactivation of bacteriophage by alkylating agents derives from reaction with the deoxyribonucleic acid moiety, probably leading to a failure of the injection process.

scholarly journals Convenient Genetic Encoding of Phenylalanine Derivatives through Their α-Keto Acid Precursors

Biomolecules ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1358
Author(s):  
Li Liu ◽  
Bohao Wang ◽  
Sheng Li ◽  
Fengyuan Xu ◽  
Qi He ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Code ◽  
Fluorescent Proteins ◽  
Coli Strain ◽  
Keto Acid ◽  
Phenylpyruvic Acid ◽  
Genetic Encoding ◽  
Keto Acids ◽  
Genetic Code Expansion ◽  
And Function

The activity and function of proteins can be improved by incorporation of non-canonical amino acids (ncAAs). To avoid the tedious synthesis of a large number of chiral phenylalanine derivatives, we synthesized the corresponding phenylpyruvic acid precursors. Escherichia coli strain DH10B and strain C321.ΔA.expΔPBAD were selected as hosts for phenylpyruvic acid bioconversion and genetic code expansion using the MmPylRS/pyltRNACUA system. The concentrations of keto acids, PLP and amino donors were optimized in the process. Eight keto acids that can be biotransformed and their coupled genetic code expansions were identified. Finally, the genetic encoded ncAAs were tested for incorporation into fluorescent proteins with keto acids.

scholarly journals The control of ribonucleic acid synthesis in bacteria. Polymerization rates for ribonucleic acids in amino acid-starved relaxed and stringent auxotrophs of Escherichia coli

1972 ◽  
Vol 128 (5) ◽  
pp. 1021-1031 ◽  
Author(s):  
W. J. H. Gray ◽  
T. G. Vickers ◽  
J. E. M. Midgley
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Normal Growth ◽  
Acid Synthesis ◽  
Amino Acid Deprivation ◽  
Ribonucleic Acids ◽  
Rate Of Polymerization ◽  
Chain Polymerization ◽  
Multiple Amino Acid ◽  
Nucleotide Bases

Polymerization rates of newly formed chains of various RNA fractions were measured in Escherichia coli CP78 (RCstr) and CP79 (RCrel) multiple amino acid auxotrophs, deprived of four amino acids essential for growth. Immediately after the onset of severe amino acid deprivation, in RCstr strains the rate of labelling of RNA by exogenous nucleotide bases was greatly diminished. At first, the initiation of new RNA chains declined faster than the rate of polymerization in RCstr organisms, but as starvation proceeded the rate of polymerization was eventually lowered to about 10% of that found during normal growth. In strain CP79 (RCrel), on the other hand, chain-polymerization rates were unaffected by amino acid withdrawal. Artificial depletion of the intracellular purine nucleotide pools in RCstr or RCrel strains by trimethoprim, before the onset of amino acid deprivation, showed that in the RCstr, but not the RCrel strain, amino acid withdrawal gave rise to an inability of the cells to utilize exogenously supplied purine or pyrimidine bases for RNA synthesis. During a prolonged starvation, the observed 100-fold decrease in the total rate of incorporation of exogenous nucleotide bases into the RNA of RCstr organisms was ascribed to a combination of a tenfold decrease in the overall rate of RNA chain polymerization, at least a fivefold decrease in the ability of the cells to utilize exogenous bases and a preferential inhibition of initiation of stable RNA chains. None of these changes occurred in the corresponding RCrel strain.

scholarly journals The control of ribonucleic acid synthesis in bacteria. The synthesis and stability of ribonucleic acids in relaxed and stringent amino acid auxotrophs of Escherichia coli

1972 ◽  
Vol 128 (5) ◽  
pp. 1007-1020 ◽  
Author(s):  
W. J. H. Gray ◽  
J. E. M. Midgley
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Heterogeneous Material ◽  
Acid Synthesis ◽  
Sucrose Density Gradient ◽  
23S Rrna ◽  
Ribonucleic Acids ◽  
Mrna Content ◽  
High Concentrations ◽  
The Stability

The biosynthesis and stability of various RNA fractions was studied in RCstr and RCrel multiple amino acid auxotrophs of Escherichia coli. In conditions of amino acid deprivation, RCstr mutants were labelled with exogenous nucleotide bases at less than 1% of the rate found in cultures growing normally in supplemented media. Studies by DNA–RNA hybridization and by other methods showed that, during a period of amino acid withdrawal, not more than 60–70% of the labelled RNA formed in RCstr mutants had the characteristics of mRNA. Evidence was obtained for some degradation of newly formed 16S and 23S rRNA species to heterogeneous material of lower molecular weight. This led to overestimations of the mRNA content of rapidly labelled RNA from such methods as simple examination of sucrose-density-gradient profiles. In RCrel strains the absolute and relative rates of synthesis of the various RNA fractions were not greatly affected. However, the stability of about half of the mRNA fraction was increased in RCrel strains during amino acid starvation, giving kinetics of mRNA labelling and turnover that were identical with those found in either RCstr or RCrel strains inhibited by high concentrations of chloramphenicol. Coincidence hybridization techniques showed that the mRNA content of amino acid-starved RCstr auxotrophs was unchanged from that found in normally growing cells. In contrast, RCrel strains deprived of amino acids increased their mRNA content about threefold. In such cultures the mRNA content of accumulating newly formed RNA was a constant 16% by wt.

scholarly journals The control of ribonucleic acid synthesis in bacteria. The synthesis and stability of ribonucleic acid in rifampicin-inhibited cultures of Escherichia coli

1971 ◽  
Vol 122 (2) ◽  
pp. 161-169 ◽  
Author(s):  
W. J. H. Gray ◽  
J. E. M. Midgley
Keyword(s):  
Escherichia Coli ◽  
Ribonucleic Acid ◽  
Acid Synthesis ◽  
Polymerization Rate ◽  
23S Rrna ◽  
Considerable Proportion ◽  
Bacterial Growth Rate ◽  
Ribonucleic Acids ◽  
E Coli ◽  
Kinetics Of

A study was made of the kinetics of labelling of the stable ribonucleic acids (rRNA+tRNA) and the unstable mRNA fraction in cultures of Escherichia coli M.R.E.600, inhibited by the addition of 0.1g of rifampicin/l. Labelling was carried out by adding either [2-14C]- or [5-3H]-uracil as an exogenous precursor of the cellular nucleic acids. From studies using DNA RNA hybridization, the kinetics of the synthesis and degradation of mRNA was followed in the inhibited cultures. Although a considerable proportion of the mRNA labelled in the presence of rifampicin decayed to non-hybridizable products, about 25% was stabilized beyond the point where protein synthesis had finally ceased. It therefore seems unwise to extrapolate the results of studies on mRNA stability in rifampicin-inhibited cultures to the situation existing in the rate of steady growth, where there appears to be little, if any, stable messenger. The kinetics of labelling of RNA in inhibited cultures indicated that the clapsed time from the addition of rifampicin to the point at which radioactivity no longer enters the total cellular ribonucleic acids is a measure of the time required to polymerize a molecule of rRNA. At 37°C, in culture grown in broth, glucose–salts or lactate salts media, exogenous [2-14C]uracil entered rifampicin-inhibited cells and was incorporated into RNA for 2 3min after the antibiotic was added. Taking this time as that required to polymerize a complete chain of 23S rRNA, the polymerization rate of this fraction in the three media was 25, 22 and 19 nucleotides added/s to the growing chains. Similar experiments in cultures previously inhibited by 0.2g of chloramphenicol/l showed virtually identical behaviour. This confirmed the work of Midgley & Gray (1971), who, by a different approach, showed that the polymerization rate of rRNA in steadily growing and chloramphenicol-inhibited cultures of E. coli at 37°C was essentially constant at about 22 nucleotides added/s. It was thus confirmed that the rate of polymerization of at least the rRNA fraction in E. coli is virtually unaffected by the nature of the growth medium and therefore by bacterial growth rate.

scholarly journals Bacteriological pollution indicators in Ogun River flowing through Abeokuta Metropolis

2017 ◽  
Vol 36 (3) ◽  
pp. 54-63
Author(s):  
S. A. Balogun ◽  
A. S. Ayangbenro ◽  
G. L. Ogunsanya ◽  
A. A. Azeez ◽  
C. O. Muonaka ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heterotrophic Bacteria ◽  
Coli Strain ◽  
World Health ◽  
Plate Count ◽  
Most Probable Number ◽  
Probable Number ◽  
Significant Difference ◽  
The Impact ◽  
Ogun River

Water resources are significant part of integrated community development policy and good health. Hence, the need to reduce the impact of natural and anthropogenic pollution causes so as to enhance water quality. The bacteriological quality of the Ogun River was investigated to determine the sanitary conditions of the water body between March and August, 2014. Total heterotrophic bacteria counts (THBC), total coliform counts (TCC) and total Escherichia coli counts (TEC) using standard plate count and Most Probable Number (MPN) techniques were determined. The isolates were identified using 16SrRNA gene. Total heterotrophic bacteria counts varied between 1.13 × 106 and 4.1 × 107 CFU/ml, TCC ranged between 2.5 × 105 and 2.33× 107 CFU/ml and TEC was between 5 × 104 and 1.3 × 106 CFU/ml. Most Probable Number of coliforms in all samples varied between 120 and 1600 MPN/100 ml. Isolated microorganisms include Escherichia coli strain SUS9EC, Escherichia coli O157:H7 strain SSI7, Escherichia coli strain BW25113, Escherichia coli strain C-X1B, and Klebsiella oxytoca strain KU-5. One-wayanalysis of variance showed significant difference within the samples at (P<0.05).The results revealed high bacteria counts which is higher than the recommended value of 1.2 × 102 for THBC, a zero E. coli count and not more than 10 coliforms per 100 ml by World Health Organization standards for drinking water.Keywords: Water, bacteria, Escherichia coli, pollution

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