The study of variability and strain selection inStreptomyces atroolivaceus

Thin-layer chromatography of extracts of submerged cultures ofStreptomyces atroolivaceus yielded, apart from mithramycin and chromocyclomycin, eleven biologically inactive metabolites. According to the spectrum of these products, the fifteen tested strains were divided into five metabolic groups. The experimental results provided a basis for discussing the relationship between the biosynthesis of mithramycin, chromocyclomycin and other metabolites.     

The study of variability and strain selection inStreptomyces atroolivaceus

A seven-step selection procedure (repeated UV irradiation, single-step application of nitrous acid, and natural selection, following each mutagenic treatment) made it possible to increase production of mithramycin byStreptomyces atroolivaceus from 40–50 μg/ml to 680–830 μg/ml,i.e. roughly by 15 to 19-fold. The UV radiation was more effective when applying lower doses, yielding about 5% survival, 2.3% survival was obtained after the treatment with nitrous acid.N-Methyl-N′-nitro-N-nitrosoguanidine applied in a buffer pH 9.0) at doses yielding more than 99% killing was less effective than the two former mutagens.     

Spontaneous and induced variability inStreptomyces nogalater producing nogalamycin

Variability in the production of nogalamycin byStreptomyces nogalater var.nogalater was followed in untreated and mutagenized populations of the standard strainNRRL 3035 and its spontaneous variant K-18 using the method of agar blocks with subsequent tests under submerged conditions. In both strains the most active variants were obtained by natural selection without mutagenic treatment; in this way productivity increased by 108% after two selection steps. Treatment with UV-radiation did not yield variants with a highly increased activity. Gamma-radiation extended the variability but, at the same time, substantially increased the number of non-producing and low-producing isolates. Relatively high yields of (+)-variants were obtained after treatment with nitrous acid but their activity did not reach that observed in the most active spontaneous variants.     

Sequence-based prediction for vaccine strain selection and identification of antigenic variability in foot-and-mouth disease virus

Identifying when past exposure to an infectious disease will protect against newly emerging strains is central to understanding the spread and the severity of epidemics, but the prediction of viral cross-protection remains an important unsolved problem. For foot-and-mouth disease virus (FMDV) research in particular, improved methods for predicting this cross-protection are critical for predicting the severity of outbreaks within endemic settings where multiple serotypes and subtypes commonly co-circulate, as well as for deciding whether appropriate vaccine(s) exist and how much they could mitigate the effects of any outbreak. To identify antigenic relationships and their predictors, we used linear mixed effects models to account for variation in pairwise cross-neutralization titres using only viral sequences and structural data. We identified those substitutions in surface-exposed structural proteins that are correlates of loss of cross-reactivity. These allowed prediction of both the best vaccine match for any single virus and the breadth of coverage of new vaccine candidates from their capsid sequences as effectively as or better than serology. Sub-sequences chosen by the model-building process all contained sites that are known epitopes on other serotypes. Furthermore, for the SAT1 serotype, for which epitopes have never previously been identified, we provide strong evidence--by controlling for phylogenetic structure--for the presence of three epitopes across a panel of viruses and quantify the relative significance of some individual residues in determining cross-neutralization. Identifying and quantifying the importance of sites that predict viral strain cross-reactivity not just for single viruses but across entire serotypes can help in the design of vaccines with better targeting and broader coverage. These techniques can be generalized to any infectious agents where cross-reactivity assays have been carried out. As the parameterization uses pre-existing datasets, this approach quickly and cheaply increases both our understanding of antigenic relationships and our power to control disease.     

The pathway ofD-cycloserine biosynthesis inStreptomyces garyphalus

Incubation experiments using washed cells and toluene treated cells ofStreptomyces garyphalus showed that O-acetyl-L-serine and hydroxyurea are intermediates in the biosynthesis ofD-cycloserine. The formation of [¹⁴C]O-ureidoserine from O-acetyl-L-serine and hydroxyurea was demonstrated by incubating an enzyme solution with¹⁴C-labelled substrates. Desalted cell-free extract catalyzed the conversion of O-ureido-D-serine toD-cycloserine in a reaction requiring ATP and Mg²⁺. The results suggested the following pathway forD-cycloserine biosynthesis.     

Herpesvirus saimiri strain variability

Herpesvirus saimiri was isolated from 22 squirrel monkeys by cocultivation of peripheral lymphocytes with permissive owl monkey kidney monolayer cells. Comparison of virion DNA fragments produced from restriction endonuclease digestion was used as a sensitive measure of strain variability. Although all isolates contained similarities and common features, 19 of the 22 were readily distinguished. Three of the isolates, however, were indistinguishable and possibly were related epidemiologically. Distinct subtypes of H. saimiri were not evident by these criteria; Peruvian, Colombian, Guyanan, and Bolivian squirrel monkeys yielded isolates without characteristic features peculiar to the geographic region. Three of three colony-born squirrel monkeys that were tested yielded a strain of virus distinct from that obtained from the mother. In separate experiments, two of three animals chosen at random yielded a strain of virus different from that originally obtained 16 and 22 months previously; only one of the three animals examined yielded the same strain of virus 22 months after the original isolation. The degree of restriction endonuclease fragment variability among H. saimiri strains appeared to be greater than previously observed for other herpesviruses.     

The effects of multilocus balancing selection on neutral variability

We studied the effect of multilocus balancing selection on neutral nucleotide variability at linked sites by simulating a model where diallelic polymorphisms are maintained at an arbitrary number of selected loci by means of symmetric overdominance. Different combinations of alleles define different genetic backgrounds that subdivide the population and strongly affect variability. Several multilocus fitness regimes with different degrees of epistasis and gametic disequilibrium are allowed. Analytical results based on a multilocus extension of the structured coalescent predict that the expected linked neutral diversity increases exponentially with the number of selected loci and can become extremely large. Our simulation results show that although variability increases with the number of genetic backgrounds that are maintained in the population, it is reduced by random fluctuations in the frequencies of those backgrounds and does not reach high levels even in very large populations. We also show that previous results on balancing selection in single-locus systems do not extend to the multilocus scenario in a straightforward way. Different patterns of linkage disequilibrium and of the frequency spectrum of neutral mutations are expected under different degrees of epistasis. Interestingly, the power to detect balancing selection using deviations from a neutral distribution of allele frequencies seems to be diminished under the fitness regime that leads to the largest increase of variability over the neutral case. This and other results are discussed in the light of data from the Mhc.     

抑制绿脓杆菌的放线菌株SM037的筛选、鉴定及发酵条件研究

从土壤样品中分离到一株对绿脓杆菌有抑制作用的放线菌SM037,初步鉴定为黄雀链霉菌黄雀变种S.canariesvar.canaries。对SM037进行液体发酵条件优化,实验表明,最适培养基配方(g/L)为:淀粉30~40,KNO32,K2HPO40.5,MgSO40.5,NaCl 0.5,FeSO40.01。抑菌活性物质的相对效价在发酵培养96-108h时达到最高峰。     

A-factor and streptomycin biosynthesis inStreptomyces griseus

Accumulating data have shown that the metabolites with a -butyrolactone ring functions as an autoregulatory factor or a microbial hormone for the expression of various phenotypes not only in a variety ofStreptomyces spp. but also in the distantly related bacteria. A-factor, as a representative of this type of autoregulators, triggers streptomycin biosynthesis and cellular differentiation inStreptomyces griseus. A model for the A-factor regulatory cascade on the basis of recent work is as follows. At an early step in the A-factor regulatory relay, the positive A-factor signal is first received by an A-factor receptor protein that is comparable in every aspect to eukaryotic hormone receptors, and then, via one or more regulatory steps, transmitted to an A-factor-responsive protein that binds to the upstream activation sequence of thestrR gene, a regulatory gene in the streptomycin biosynthetic gene cluster. The StrR protein thus induced appears to activate the other streptomycin biosynthetic genes. This review summarizes the characteristics of A-factor as a microbial hormone and the A-factor regulatory relay leading to streptomycin production.     

Plasmid and chloramphenicol production inStreptomyces venezuelae

A strain ofStreptomyces venezuelae described as having been cured of chloramphenicol production, was mutagenised with ultraviolet light and chloramphenicol-producing clones were obtained from the surviving population. Since this suggests that the supposed cured strain has not lost the genetic capacity of chloramphenicol synthesis, alternative explanations are offered.     

Induction of pristinamycins production inStreptomyces pristinaespiralis

Summary Pristinamycins production in a mutant ofStreptomyces pristinaespiralis (strain Cl6/4) blocked in pristinamycins biosynthesis was induced by different commercial lactones on solid media. The activity of the -lactone ring molecules was improved when the length of the substituted carbon chain was increased. Pristinamycins induction was also obtained with the endogenous A aactor ofS. griseus. The minimum effective concentration of the A factor was 250 times lower than that of exogenous lactones. Addition of exogenous -butyrolactone did not affect antibiotic production by a hyperproducing mutant ofStreptomyces pristinaespiralis (strain Pr11) even in the presence of surfactant like Tween 80 or Triton X 100. Three hours before the initiation of antibiotic production, the hyperproducing strain Pr11 produced an extracellular factor extractable with ethyl acetate at pH 7 and able to induce the antibiotic production of strain Cl6/4.     

Role of ATP-glucokinase and polyphosphate glucokinase inStreptomyces aureofaciens

The activity of ATP-glucokinase and of polyphosphate glucokinase was examined during growth of the actinomyceteStreptomyces aureofaciens 8425 under conditions of intense chlortetracycline (CTC) synthesis. ATP-glucokinase was active in the strain only during the logarithmic phase of culture growth; the activity of polyphosphate glucokinase appears only at the end of the logarithmic phase of growth and rises in parallel with the rate of CTC biosynthesis in the stationary phase. During the rise of activity of polyphosphate glucokinase and of CTC biosynthesis the cells accumulate sugar phosphates, mainly glucose-6-phosphate. It appears that the biosynthesis of CTC inStreptomyces aureofaciens takes place at the expense of glycolysis, using up the high-energy phosphate of high-molecular polyphosphates.     

Isolation and characterization of glycocalyx inStreptomyces aureofaciens

The surface layer, considered to be glycocalyx according to electron-microscopic observations, was separated from a lowproduction strain ofStreptomyces aureofaciens by solubilization with urea and subsequent sonication. The isolation procedure was developed using various agents; neutral phosphatase served as a marker indicating the amount of the material released. The peripheral structure consisted predominantly of glycoprotein and differed from S-layers.     

Genetic variability under mutation selection balance

A fundamental problem in evolutionary genetics is understanding how high levels of genetic variation in quantitative traits are maintained in natural populations. Variation is removed by the natural selection of individuals with optimal phenotypes and is recovered by mutation; however, previous analyses had indicated that a mutation-selection balance was insufficient to maintain observed levels of genetic variation in these traits. Using more general models, however, it has recently been shown that it is indeed a sufficient mechanism. These models can be used to explore other phenomena in evolutionary biology.     

Production of quinomycin A inStreptomyces lasaliensis

In addition to lasalocid, an oligoether coccidiostatic compound, other compounds are synthesized byStreptomyces lasaliensis. Mutants producing either of two antibiotics, lasalocid A or quinomycin A (an antibiotic of quinoxaline character), were obtained by natural selection and by mutagenesis. Methods of isolation, purification and estimation of both compounds were established.     

Induction of β-D-Glucosidase inStreptomyces granaticolor

beta-D-glucosidase in Streptomyces granaticolor is an inducible enzyme. Methyl-beta-D-glucoside or cellobiose, added to a glycerol-containing medium, are most suitable inducers. The activity of beta-D-glucosidase in a culture fully induced by cellobiose is 50 times higher than the basal level of the enzyme. beta-D-glucosidase is an intracellular enzyme, whose inducibility differ with culture age and reaches its maximum in a 10-h-old mycelium. The enzyme synthesis begins 2 h after the addition of the induced and reaches its maximum after a 10-h-induction.     

Production of rhodomycins inStreptomyces griseoruber 4620

The strainStreptomyces griseoruber 4620 produces, besides the anthracycline antibiotics beromycins, some other anthracyclines of the rhodomycin type. Twelve isolates exhibiting a higher antibiotic activity (up to 2.5×), as compared to the parent strain, were obtained after a spontaneous selection. The following species were isolated from the hydrolysate of mycelial extract: β-rhodomycinone, β-isorhodomycinone, α₂-rhodomycinone and 10-deoxy-β-rhodomycinone, which has not yet been described.