RESUMEN
The morphological and physiological characteristics of Bacillus thuringiensis strains were analyzed and conditions for obtaining culture fluid with maximum yield of secreted RNases were determined. Zymographic analysis showed that culture fluid of B. thuringiensis strains along with low-molecular-weight (15-20 kDa) RNases contained enzymes with a molecular weight ~55 kDa and their content depended on the duration and conditions of culturing. Preparations based on B. thuringiensis culture fluid were effective against human influenza virus A/Aichi/2/68 (H3N2). In experiments on mice infected with 10 LD50 influenza virus strain A/Aichi/2/68 (H3N2), we selected effective variants of preparations based on culture fluid of B. thuringiensi strains for preventive administration that provided reliable protection of infected animals (protection coefficient 50%), close to that of the reference drug Tamiflu.
Asunto(s)
Antivirales/farmacología , Bacillus thuringiensis/efectos de los fármacos , Bacillus thuringiensis/virología , Subtipo H3N2 del Virus de la Influenza A/patogenicidad , Virus de la Influenza A/patogenicidad , Kobuvirus/patogenicidad , Oseltamivir/farmacología , Humanos , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Virus de la Influenza A/efectos de los fármacos , Gripe Humana/microbiología , Kobuvirus/efectos de los fármacosRESUMEN
The results of studies of a newly isolated Serratia species K-57 strain are presented. The strain is characterized by antiviral activity towards human influenza A/Aichi/2/68/H3N2, vaccinia, mouse smallpox, and herpes simplex-2 viruses. The detected characteristics of the strain, including the data on activities on nucleolytic enzymes, recommend it for the development of therapeutic and preventive antiviral drugs.
Asunto(s)
Antivirales/farmacología , Proteínas Bacterianas/farmacología , Desoxirribonucleasas/farmacología , Ribonucleasas/farmacología , Serratia/química , Animales , Antivirales/aislamiento & purificación , Proteínas Bacterianas/aislamiento & purificación , Chlorocebus aethiops , Desoxirribonucleasas/aislamiento & purificación , Perros , Herpesvirus Humano 2/efectos de los fármacos , Herpesvirus Humano 2/crecimiento & desarrollo , Humanos , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Subtipo H3N2 del Virus de la Influenza A/crecimiento & desarrollo , Células de Riñón Canino Madin Darby , Ratones , Pruebas de Sensibilidad Microbiana , Ribonucleasas/aislamiento & purificación , Virus Vaccinia/efectos de los fármacos , Virus Vaccinia/crecimiento & desarrollo , Virus de la Viruela/efectos de los fármacos , Virus de la Viruela/crecimiento & desarrollo , Células VeroRESUMEN
Spectra of amplitudes of polarization of erythrocytes, yeast cells, and latex particles in the range of 1-10 Hz were investigated by the method of dielectrophoresis. Positive dielectrophoresis of erythrocytes and yeast cells the frequency range of 60 - 100 Hz was revealed. The theoretically positive dielectrophoresis was evidenced by the occurrence of channels across the cell membrane and bacterial cell wall.
Asunto(s)
Bacterias/química , Membrana Celular/química , Polaridad Celular , Pared Celular/química , Eritrocitos/química , Levaduras/química , Animales , Humanos , MicroesferasRESUMEN
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.
Asunto(s)
Sedimentos Geológicos/microbiología , Methylobacteriaceae/clasificación , ADN Bacteriano/análisis , Agua Dulce , Filogenia , SiberiaRESUMEN
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.
Asunto(s)
Bacterias/genética , Bacterias/ultraestructura , ADN Bacteriano/genética , Agua Dulce , Bacterias/clasificación , Composición de Base , Sedimentos Geológicos/microbiología , Siberia , Microbiología del AguaRESUMEN
The process of isolation of highly purified human lymphotoxin was studied and optimized. A set of methods providing an increase in the content of the target product in the biomass (plasmid DNA amplification and selection of clones of transformed cells) was applied at the stage of cultivation. A two-step purification scheme (ion-exchange chromatography on DEAE-Biotone A1 and hydroxylapatite) was developed. A number of physical characteristics were studied.
Asunto(s)
Linfotoxina-alfa/genética , Cromatografía por Intercambio Iónico , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Escherichia coli/genética , Humanos , Linfotoxina-alfa/aislamiento & purificación , Linfotoxina-alfa/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismoRESUMEN
Study of the biosynthesis of human alpha 2-interferon and tumor necrosis factor in derivatives of E. coli SG200-50 recipient strain containing pIF16, pTNF311, and pLT21 plasmids demonstrated elimination of the plasmids from the cells without degradation thereof in the course of culturing. Methods increasing the content of the end products in the biomass are proposed.
Asunto(s)
Interferón Tipo I/biosíntesis , Linfotoxina-alfa/biosíntesis , Factor de Necrosis Tumoral alfa/biosíntesis , Farmacorresistencia Microbiana , Escherichia coli/genética , Escherichia coli/metabolismo , Marcadores Genéticos , Humanos , Plásmidos , Proteínas Recombinantes/biosíntesisRESUMEN
Bme 361 I, a new site-specific type II deoxyribonuclease, was purified from Bacillus megaterium 361 by chromatography on phosphocellulose P 11 and hydroxylapatite. The enzyme recognizes and cleaves the nucleotide sequence 5'-GG decreases CC-3' in double-strand DNA. Thus it is a true isoschizomer of deoxyribonucleases Hae III and BspR I.
Asunto(s)
Bacillus megaterium/enzimología , Desoxirribonucleasas/metabolismo , Autorradiografía , Bacteriófago lambda/metabolismo , Cromatografía Liquida , ADN Bacteriano/metabolismo , ADN Viral/metabolismo , Electroforesis en Gel de Poliacrilamida , Hidrólisis , Plásmidos , Especificidad por SustratoRESUMEN
The method for analysis of microorganisms for the presence of the modification-restriction systems has been developed. The method has permitted to detect more than 10 new producing strains of restrictases including microorganisms of Rhizobium genus. Some of them are promising for practical use. It has been shown that using selection of clones the strain productivity can be increased. The purification process for the majority of restrictases has been proposed. Some physical and catalytic properties of new enzymes have been studied.
Asunto(s)
Bacterias/enzimología , Enzimas de Restricción-Modificación del ADN/biosíntesis , Enzimas de Restricción-Modificación del ADN/aislamiento & purificación , Técnicas Bacteriológicas , Bacteriófago lambda , Fenómenos Químicos , Química Física , ADN Viral/metabolismo , Hidrólisis , Mapeo Restrictivo , Especificidad por SustratoRESUMEN
A new site-specific restriction endonuclease Rme211 from Rhizobium meliloti has been shown to recognize the following nucleotide sequence 5'-ATCGAT-3' in the double-stranded DNA. Thus, the enzyme is a true isoschizomer for restriction endonucleases Bsu151 and ClaI.
Asunto(s)
Enzimas de Restricción del ADN/metabolismo , Desoxirribonucleasas de Localización Especificada Tipo II/metabolismo , Rhizobium/enzimología , Bacteriófago lambda/metabolismo , Cromatografía DEAE-Celulosa , Enzimas de Restricción del ADN/aislamiento & purificación , ADN Viral/análisis , Desoxirribonucleasas de Localización Especificada Tipo II/aislamiento & purificación , Especificidad por SustratoAsunto(s)
Desoxirribonucleasas de Localización Especificada Tipo II/metabolismo , Klebsiella pneumoniae/enzimología , Bacteriófago lambda/metabolismo , Secuencia de Bases , ADN Viral/metabolismo , Desoxirribonucleasas de Localización Especificada Tipo II/antagonistas & inhibidores , Hidrólisis , Datos de Secuencia Molecular , Cloruro de Potasio , Cloruro de SodioRESUMEN
The 99 base pair poly(dA).poly(dT) stretch was inserted into EcoRI site of pBR plasmid. The effect of this insertion on tet-gene expression from P2 promoter, and bla-gene expression from P1 and P3 promoters was studied. The insertion results in a slight enhancement of the resistance of the cells to tetracycline and a decreased sensitivity to ampicillin. It is suggested that these findings reflect the effect of insertion on the transcription of the corresponding genes.
Asunto(s)
Desoxirribonucleasa EcoRI , Regulación de la Expresión Génica , Poli dA-dT , Polidesoxirribonucleótidos , Resistencia a la Ampicilina/genética , Secuencia de Bases , ADN/genética , Electroforesis en Gel de Agar , Plásmidos , Regiones Promotoras Genéticas , Resistencia a la Tetraciclina/genética , Transformación GenéticaAsunto(s)
Ética , Relaciones Familiares , Relaciones Interpersonales , Matrimonio , Conducta Sexual , Cambio Social , Derechos de la Mujer , Américas , Conducta , Países Desarrollados , Economía , Europa (Continente) , Composición Familiar , América del Norte , Educación Sexual , Factores Socioeconómicos , U.R.S.S. , Estados UnidosRESUMEN
The effect of some adenyl precursors of DNA synthesis on the mutagenic activity of hydroxylamine (HA) is studied. It is shown that the addition of adenine to a suspension of Escherichia coli B cells increases the yield of mutants by more than two times as compared with HA alone. The effects of adenosine, AMP and dAMP are somewhat different. It is suggested that the increase of the HA mutagenic effect produced by the addition of adenine may be due to: 1) the excess of the amount of adenylic precursors of DNA synthesis over guanilic ones, which promotes the erroneous base-pairing during the replication of the HA modified template; 2) the modification of adenylic precursors by HA into N6-oxy-dATP, and their incorporation into DNA. The mutagenic effect of N6-hydroxyadenosine, the product of the adenine modification by HA, in E. coli B pur- was studied. The experiments showed that N6-hyrdoxyadenosine induced about 1% of mutations, a relatively low lethal effect (the cell survival was 80%), and provided a high mutagenic action of this compound.
Asunto(s)
Adenina/farmacología , Adenosina Monofosfato/farmacología , Adenosina/farmacología , Desoxiadenosinas/farmacología , Hidroxilaminas/farmacología , Mutágenos , Interacciones Farmacológicas , Escherichia coli/efectos de los fármacosRESUMEN
The effect of some adenyl precursors of DNA synthesis on the mutagenic activity of hydroxylamine (HA) is studied. It is shown that the addition of adenine to a suspension of Escherichia coli B cells increases the yield of mutants by more than two times as compared with HA alone. The effects of adenosine, AMP and dAMP are somewhat different. It is suggested that the increase of the HA mutagenic effect produced by the addition of adenine may be due to: 1) the excess of the amount of adenylic precursors of DNA synthesis over guanilic ones, which promotes the erroneous base-pairing during the replication of the HA modified template; 2) the modification of adenylic precursors by HA into N6-oxy-dATP, and their incorporation into DNA. The mutagenic effect of N6-hydroxyadenosine, the product of the adenine modification by HA, in E. coli B pur- was studied. The experiments showed that N6-hydroxyadenosine induced about 1% of mutations, a relatively low lethal effect (the cell survival was 80%), and provided a high mutagenic action of this compound.
Asunto(s)
Nucleótidos de Adenina/farmacología , Adenina/farmacología , Adenosina/farmacología , Hidroxilaminas/farmacología , Mutágenos , Adenosina/análogos & derivados , Interacciones Farmacológicas , Escherichia coli/efectos de los fármacosRESUMEN
The effect of some adenyl precurosors of DNA synthesis on the mutagenic activity oh hydroxylamine (HA) is studied. It is shown that the addition of adenine to a suspension of Escherichia coli Bcells increases the yield of mutants by more than two times as compared with HA alone. The effects of adenosine, AMP and dAMP are somewhat different. It is suggested that the increase of the HAmutagenic effect produced by the addition of adenine may be due to:1) the excess of the amount of adenylic precursors of DNA synthesis over guanilic ones, which promotes the erroneous base-pairing during the replication of the HA modified template; 2) the modification of adenylic precursors by HA into N6-oxy-dATP, and their incorporation into DNA. The mutagenic effect of N6-hydroxyadenosine, the product of the adenine modification by HA, in E. coli Bpur- was studied. The experiments showed that N6-hydroxyadenosine induced about 1% of mutations, a relatively low lethal effect (the cell survival was 80%), and provided a high mutagenic action of this compound.