RESUMEN
Analysis of molecular-genetic polymorphism of barley varieties was performed using AFLP-method. A system for identification and differentiation of barley varieties based on AFLP-markers was developed. Results of testing for 19 varieties indicate a high differential ability of the developed system. Identification of varieties can be conducted using one of two proposed discriminatory sets of AFLP-markers. Based on the calculated genetic distances a dendrogram of phylogenetic relations between the varieties was constructed. The dendrogram revealed a separated origin of varieties of malting and feed directions.
Asunto(s)
ADN de Plantas/análisis , Hordeum/genética , Filogenia , Polimorfismo Genético , Selección Genética , Alelos , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Alimentación Animal , Cerveza , Cartilla de ADN/biosíntesis , ADN de Plantas/clasificación , ADN de Plantas/genética , Frecuencia de los Genes , Marcadores Genéticos , Hordeum/clasificación , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
In this work we have demonstrated two independent real-time PCR methods for detection single nucleotide polymorphism (SNP) of genes csn and acyl-CoA: diacylglycerol acyltransferase 1 (dgat) in cattle. We have analyzed 296 samples of milk production cattle of Ukrainian breeding. The genotype frequencies were AA--0.58, AB--0.34, BB--0.08 for csn gene and for dgat gene--AA--0.7, AK--0.26, KK--0.04. High efficiency of so called "anti-primer" method was shown. Duration of anti-primer PCR reaction was about 2-2.5 hours only and provided full investigation of unknown gene allele.
Asunto(s)
Caseínas/genética , Cartilla de ADN , Diacilglicerol O-Acetiltransferasa/genética , Lactancia/genética , Polimorfismo de Nucleótido Simple , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Alelos , Animales , Bovinos , Industria Lechera , Femenino , Frecuencia de los Genes , UcraniaRESUMEN
The WHO approved protocol of pandemic influenza virus A/H1N1 identification by the method of one-step RT-PCR was adopted. The cost of research was decreased due to adoption of RNA purification method and utilizing of the common enzyme systems for RT-PCR.
Asunto(s)
Brotes de Enfermedades , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Diagnóstico Diferencial , Humanos , Subtipo H1N1 del Virus de la Influenza A/genética , Gripe Humana/epidemiología , Mucosa Bucal/virología , Sensibilidad y Especificidad , UcraniaRESUMEN
Bacteria with high N2-fixing activity were isolated from the root zone of spring wheat grown on leach chernozem and soddy podzolic soil in Ukrainian marshy woodlands. They were characterized by phenotypic signs and investigated with the help of molecular-genetic methods. On the basis of diagnostic signs the investigated strains were referred to Azospirillum brasilense from Azospirillum genus. Their 3'- and 5'-thermal 16S RNA hypervariable sites with length from 373 to 395 nucleotides were amplified and sequenced. The comparative analysis of results confirmed the 100% identity of 16S RNA sequences from investigated bacteria with the same sequences of A. brasilense from Gene Bank database. Thus the results of sequence analysis agree with results obtained during the investigation of phenotypic signs.
Asunto(s)
Azospirillum/clasificación , Azospirillum/aislamiento & purificación , Raíces de Plantas/microbiología , Triticum/microbiología , Azospirillum/genética , ADN de Plantas/genética , Electroforesis en Gel de Agar , Raíces de Plantas/crecimiento & desarrollo , Reacción en Cadena de la Polimerasa , ARN de Planta/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Triticum/crecimiento & desarrollo , UcraniaRESUMEN
The influence of simulated microgravitation (clinostating) on the proceeding of mixed virus infection in Zhuravushka variety potato plants, infected with potato viruses X and M (PVX, PVM) was investigated. It was shown, that sensitivity of "virus-plant" system to microgravitation depends both on pathogene species and clinostating mode. Vertical clinostating was found to be more effective than horizontal one. By means of biotest, PCR and ELISA it was detected that potato plants, infected with PVX and PVM are released from PVX under the influence of clinostating during 19-47 days. Assume, that microgravitation is able to induce some protective reactions in plants that influence virus reproduction.
Asunto(s)
Enfermedades de las Plantas/virología , Virus de Plantas/aislamiento & purificación , Solanum tuberosum/crecimiento & desarrollo , Solanum tuberosum/virología , Simulación de Ingravidez , Electroforesis en Gel de Agar , Genes Virales/genética , Microscopía Electrónica , Enfermedades de las Plantas/etiología , Virus de Plantas/crecimiento & desarrollo , Virus de Plantas/ultraestructura , Reacción en Cadena de la Polimerasa , RotaciónRESUMEN
The chemical synthesis of a modified gene fragment that codes IgG-binding domain of protein G of Streptococcus sp. (SPG) was done. Two copies of gene fragmentes SPG were cloned into plasmid vector pET-24(a), for expression of recombinant truncated protein G (rSPG) in the culture of Escherichia coli. IgG-binding capacity and specificity for rSPG was confirmed by model assay such interaction in vitro. Basing on the obtained data we affirm that truncated recombinant protein G is able to bind immunoglobulin G in mammalians and can be used for purification of IgG from sera by affinity chromatography or for synthesis of immunoassay conjugates with broad range specificity.
Asunto(s)
Proteínas Bacterianas , Técnicas para Inmunoenzimas/métodos , Inmunoglobulina G/metabolismo , Proteínas Recombinantes , Streptococcus/metabolismo , Proteínas Bacterianas/biosíntesis , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Escherichia coli/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Streptococcus/genéticaRESUMEN
We have carried out comparative studies of double-stranded RNA (dsRNA) of viral nature isolated from sugar beet leaves and from mycelium of microscopic fungi using different methods such as PAAG electrophoresis and by polymerase chain reaction (PCR). It was shown that the fragments of dsRNA from sugar beet leaves and from mycelium microscopic fungi had the identical electrophoretic pattern and the same size (1.8 and 2.0 kbp). Using PCR technique it was shown, that isolated dsRNA have a common template for amplification. Electron microscopy of PCR-positive mycelium allows us to detect the virus particles of the spherical form with diameter 30-40 nm. The obtained data confirm our previous suppositions, concerning the belonging of isolated dsRNAs (size 1.8 and 2.0 kbp) to new mycovirus targeted a microscopic fungus, instead of beet cryptic viruses.
Asunto(s)
Basidiomycota/virología , Beta vulgaris/microbiología , Virus de Plantas/aislamiento & purificación , ARN Bicatenario/análisis , ARN Viral/análisis , Basidiomycota/ultraestructura , Beta vulgaris/virología , Electroforesis en Gel de Poliacrilamida , Microscopía Electrónica , Micelio/ultraestructura , Micelio/virología , Reacción en Cadena de la PolimerasaRESUMEN
Species composition of fungi, isolated from the sugar beet leaves, roots and rhizosphere, collected in Poltava and Kyiv regions has been studied. Species Alternaria, Trichoderma, Cladosporium, Acremonium, Penicillium, Fusarium, Mycelia sterilia were found in all investigated samples. Basidiomycetes were isolated from the leaf samples in separate cases.
Asunto(s)
Beta vulgaris/microbiología , Hongos Mitospóricos/aislamiento & purificación , Hojas de la Planta/microbiología , Raíces de Plantas/microbiología , Acremonium/aislamiento & purificación , Alternaria/aislamiento & purificación , Basidiomycota/aislamiento & purificación , Cladosporium/aislamiento & purificación , Fusarium/aislamiento & purificación , Penicillium/aislamiento & purificación , Trichoderma/aislamiento & purificación , UcraniaRESUMEN
We have developed diagnostic system for revealing 35S promoter CaMV and transgene CryIIIA in transgenic potato NL Russet Burbank and NL Superior companies Monsanto. Using the data about nucleotide sequences of 35S promoter (AF234316 GenBank) and gene CryIIIA (X70979 GenBank) oligonucleotides primers were synthesized for polymerase chain reaction (PCR). The technique of genomic DNA isolation from plant material, suitable for PCR has been optimized. Using Touchdown PCR method presence of specific transgene in a Bt-potato was shown. This method can be used in routine laboratory practice for revealing 35S promoter in genetically modified plants and for identification of specific transgene CryIIIA in the transformed Bt-plants.