ABSTRACT
Permafrost is widely distributed in the Northern Hemisphere, and its age reaches hundreds of thousands and millions of years. Permafrost contains alive microorganisms which are not frozen due to relatively high temperature of the environment (-2...-8 degrees C), but the microorganisms are immobilized and therefore aged probably similar to the age of permafrost. Longevity of the relict microbial cells is related obviously to their mechanism of protection against heat, radiation, free radicals and other damaging agents. A strain of Bacillus sp. was isolated from permafrost aged of about 3 million years, 16S rDNA sequence was identified and preliminary testing of bacterial culture on Drosophila melanogaster and mice was made. Immune stimulation and improvement of physical condition were observed, and that, together with the age of the microbial cells, presents the relict microorganisms as objects of gerontology.
Subject(s)
Aging/immunology , Aging/physiology , Bacillus/growth & development , Bacillus/physiology , Cold Climate , Geriatrics/methods , Animals , Bacillus/isolation & purification , DNA, Bacterial/genetics , Drosophila melanogaster/growth & development , Drosophila melanogaster/immunology , Drosophila melanogaster/microbiology , Immunity, Cellular/physiology , Immunity, Humoral/physiology , Mice , Mice, Inbred Strains , RNA, Ribosomal, 16S/genetics , Siberia , Soil Microbiology , Spleen/immunology , Thymus Gland/immunology , Time FactorsABSTRACT
A strain producing a restriction endonuclease was isolated from soil samples and identified as the Arthrobacter sp. strain Ck256. The enzyme produced by this strain was termed Asi2561. The isolation procedure for this enzyme was described, and the optimal conditions for its function were determined. It was shown that the restriction endonuclease Asi256I is a true isoschizomer of MboI, it has a temperature optimum of 6 degrees C, and can be used in molecular-biological and genetic-engineering studies performed at low temperatures.
Subject(s)
Arthrobacter/enzymology , DNA Restriction Enzymes/isolation & purification , DNA Restriction Enzymes/metabolism , Arthrobacter/classification , Arthrobacter/isolation & purification , Bacteriophage lambda/genetics , Cold Temperature , DNA Restriction Enzymes/classification , DNA, Viral/genetics , DNA, Viral/metabolism , Soil Microbiology , Substrate SpecificityABSTRACT
Microbiological analysis of samples of sedimentary rocks from various eras of the geological history of the Baikal rift has enabled us to isolate a large number of microorganisms that can be classified into new, previously undescribed species. The present work deals with the identification and study of the morphological, biochemical, and physiological properties of one such strain, Che 82, isolated from sample C-29 of 3.4-3.5 Ma-old sedimentary rocks taken at a drilling depth of 146.74 m. As a result of our investigations, strain Che 82 is described as a new bacterial species, Roseomonas baikalica sp. nov., belonging to the genus Roseomonas within the family Methylobacteriaceae, class Alphaproteobacteria.
Subject(s)
Geologic Sediments/microbiology , Methylobacteriaceae/classification , DNA, Bacterial/analysis , Fresh Water , Phylogeny , SiberiaABSTRACT
Oil-oxidizing microorganisms have been sampled in various regions of Siberia and used in strain associations, which degrade n-alkanes of oil from various fields by 64-92% after 6 days of growth in a wide temperature range. These strains are salt-tolerant and psychrotolerant. They are compatible with aboriginal soil microflora. Promising results have been obtained in experiments on growing plants on oil-polluted soil purified with a biodegrader of this series.
Subject(s)
Alkanes/metabolism , Mineral Oil/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Cold Temperature , SiberiaABSTRACT
Genetic and biochemical methods and morphological examination were used to study microorganisms isolated from samples of deep drilling of the Lake Baikal bottom sediments. Based on blot hybridization patterns, the strains investigated were divided into several groups according to the degree of homology of their genomic DNA. Morphological, biochemical, and ultrastructural characteristics of bacterial strains are described, and their compliance with the genomic analysis data is demonstrated.
Subject(s)
Bacteria/genetics , Bacteria/ultrastructure , DNA, Bacterial/genetics , Fresh Water , Bacteria/classification , Base Composition , Geologic Sediments/microbiology , Siberia , Water MicrobiologyABSTRACT
The method of the cultivation of M .tuberculosis in cell subcultures was tested. Five consecutive passages of 2 M. tuberculosis strains were carried out with similar inoculation and cultivation conditions. Mycobacterial cells preserved their morphological characteristics in the course of all passages. The method of M. tuberculosis cultivation in subcultured cells made it possible to study of the physiology of M. tuberculosis under conditions, most approximated to the natural tuberculosis infection in humans.
Subject(s)
Mycobacterium tuberculosis/growth & development , Animals , Chlorocebus aethiops , Culture Media , Time Factors , Vero CellsABSTRACT
A total of 115 collection strains of Bacillus thuringiensis, belonging to various subspecies, have been studied for the presence of DNA restriction-modification systems. Restriction endonucleases of 13 strains have been isolated and characterized. No considerable correlations between the taxonomic positions of the bacteria and the specificities of the endonucleases isolated have been detected. It is concluded that the enzymes with identical specificities are present in both the crystalliferous and acrystalliferous strains of the same subspecies.
Subject(s)
Bacillus thuringiensis/enzymology , DNA Restriction Enzymes/metabolism , DNA Restriction Enzymes/biosynthesis , DNA Restriction Enzymes/isolation & purification , Substrate SpecificityABSTRACT
A new method of testing restriction nucleases is proposed. This method is based on high-temperature treatment of crude cell extracts. Disrupted cells were heated at 50-60 degrees C, centrifuged, and assayed for restrictases. This method provides the opportunity for screening new enzymes in microbial strains enriched with nonspecific restrictases. High-temperature treatment of cell extracts of certain producers reduces the number of steps of the procedure used for isolating high-purity restrictases; the resulting preparations are capable of maintaining high enzymatic activity during long-term storage. It was shown that high-temperature treatment can be applied not only to thermophilic but also to mesophilic strains of microorganisms of different taxa.
Subject(s)
Bacteria/enzymology , DNA Restriction Enzymes/isolation & purification , Biotechnology/methods , Hot Temperature , Quality ControlABSTRACT
Genomic DNA of the entomopathogenic bacterium Bacillus thuringiensis was analyzed by the genomic fingerprinting technique. The biotin-labeled single-stranded DNA of the phage M13 was used as a marker of hypervariable sequences. A procedure for analyzing the differentiation among various Bacillus thuringiensis strains was developed. Characteristic patterns of fingerprints were obtained for several strains, the main representatives of subspecies that are most frequently used in the manufacture of bacterial insecticides, such as subsp. thuringiensis, subsp. kurstaki, and subsp. galleriae. Because no essential differences were revealed in band patterns upon comparing fingerprints of crystal-producing bacterial strains with those of acrystallic mutants, it was assumed that the loss of crystal-producing ability in the insect pathogen Bacillus thuringiensis is not connected with significant rearrangement of its genome.
Subject(s)
Bacillus thuringiensis/isolation & purification , Bacteriophage M13/genetics , Biotin/metabolism , DNA Fingerprinting , DNA, Viral/genetics , Bacillus thuringiensis/genetics , DNA, Viral/metabolism , PhenotypeABSTRACT
HpyNI is the first restriction endonuclease which has been isolated from pathogenic Helicobacter pylori. HpyNI, a new isoschizomer of ScrFI, recognizes and cleaves the palindromic sequence 5'CCNGG-3'.
Subject(s)
Deoxyribonucleases, Type II Site-Specific/isolation & purification , Helicobacter pylori/enzymology , Gastric Mucosa/microbiology , Humans , Isoenzymes/isolation & purification , Male , Stomach Ulcer/microbiologyABSTRACT
To analyze DNA polymorphisms of various bacterial strains, a nonradioactive variant of the genomic fingerprinting method was developed. The method was based on the application of biotin-labeled single-stranded phage M13 DNA as a probe. Characteristic patterns of fingerprints obtained by MvaI, HaeIII, and HinfI restriction enzymes are presented for several species of bacilli and other bacteria. The advantages of this method in microbiology for the identification and characterization of different microbial strains are shown.
Subject(s)
Bacteria/genetics , Bacteriophage M13/genetics , DNA Fingerprinting/methods , DNA, Viral , Genome, Bacterial , Bacillus/genetics , Bacteria/classification , Biotin , DNA Probes , Escherichia coli/genetics , Vibrio/geneticsABSTRACT
Luminescent bacteria isolated from the waters of the Black Sea and of the Indian and Pacific Oceans were tested for the presence of restriction endonucleases. For 12 of 19 restriction enzyme producers revealed, restriction sites were determined. The enzymes were identified as isoschizomers of AfIII, BanI, HaeIII, PstI, Sau3AI, and Sau96I. Possible participation of the restriction and modification enzymes in the interaction of bacterial symbionts with the animal macrosymbiont is discussed.
Subject(s)
Bacteria/metabolism , DNA Restriction Enzymes/biosynthesis , Luminescent Measurements , Bacteriophage lambda/genetics , DNA, Viral/metabolism , Water MicrobiologyABSTRACT
Using the most effective rapid method for the detection of restriction endonucleases (ENases) in microorganisms, 32 thermophilic producers have been isolated. All strains belong to the genus Bacillus. Thermostable isoschizomers of ENases, such as AvaI, BbvI, BbvII, BclI, BsaBI, BsiYI, BsrI, BstEII, BstNI, Cfr10I, ClaI, FspI, HaeIII, HpaII, Ksp632I and SfeI, were isolated.
Subject(s)
Bacillus/enzymology , DNA Restriction Enzymes/metabolism , Soil Microbiology , Base Sequence , DNA Restriction Enzymes/isolation & purification , Enzyme Stability , Hot Temperature , Molecular Sequence Data , Species SpecificityABSTRACT
Restriction endonucleases have been isolated from 26 strains of thermophilic strains of the Bacillus genus, their recognition sequences were determined, and for 15 of them cleavage sites identified. The enzymes proved to be isoschizomers of known endonucleases BstNI, EarI, HaeIII, HpaII, Cfr10I, BsiYI, BclI, BbvII, BbvI, BstEII, BsaBI, BsrI, FspI, ClaI, SfeI.
Subject(s)
Bacillus/enzymology , DNA Restriction Enzymes/isolation & purification , DNA Restriction Enzymes/metabolism , Substrate SpecificitySubject(s)
Bacillus/enzymology , Deoxyribonucleases, Type II Site-Specific/isolation & purification , Isoenzymes/isolation & purification , Bacteriophage lambda/metabolism , Base Sequence , DNA, Viral/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Isoenzymes/metabolism , Molecular Sequence DataABSTRACT
Sensitivity of several express-methods used for detection of bacterial endonucleases was compared. The most sensitive method is that employing Triton X-100.