2/2013

Author

page: 4-12 pp
DOI: 10.11134/btp.2.2013.1
М.К. Иманбекова1, Е.В. Жолдыбаева1, Т.К. Есентаев2, К.Т. Момыналиев1
1Национальный центр биотехнологии, г. Астана
2Агенство РК по делам спорта и физической культуры

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Abstract

Ability, causing the ability to become an elite athlete have a genetic basis. A number of factors will determine the success of the sport: genetics, epigenetics, exercise, nutrition, motivation, achievement in the field of exercise equipment, etc. Genetics determines the important components of athletic success, such as strength, power, endurance, muscle fiber size and composition, flexibility, neuromuscular coordination, temperament and other phenotypes. Thus, the success of an athlete is largely determined by heredity about 66% of differences between athletes due to genetic factors. The remaining difference is explained by environmental factors. However, despite the obvious role of genetics in a sports results, accumulated little definitive evidence pointing to the contribution of specific genetic variants on the progress in the sport.

This can be causing of polygenic (many genes of small effect) influence of genes. This review examines the importance of genetic markers for predicting sporting success or adjustments to the training process of elite athletes.

Keywords: genetics, sports, polymorphism, endurance, strength, speed.

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Author

page: 12-16 pp
DOI: 10.11134/btp.2.2013.2
К.Х. Жуматов, М.Х. Саятов, А.И. Кыдырманов
РГП «Институт микробиологии и вирусологии» КН МОН РК, г. Алматы
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Abstract

Influenza A viruses belong to the family Orthomyxoviridae and are characterized by single-stranded RNA genome of negative polarity segmented into eight fragments coding for at least 10 proteins. Two glycoproteins on the virion surface - hemagglutinin and neuraminidase are involved in the processes of attachment to the host cell and release of viral progeny. To date, 16 subtypes of hemagglutinin and 9 - neuraminidase is known. Their various combinations determine the antigenic variant of influenza virus: H1N1 H3N2 H5N1, H7N9, etc. The review article is devoted to the actual variants of influenza A virus with hemagglutinin H7. According to clinical and pathological characteristics the low pathogenic avian influenza manifests itself in the form of respiratory and intestinal disorders, reproductive disorders, and is caused by viruses on all varieties of hemagglutinin. Highly pathogenic avian influenza, is usually caused only by representatives of subtypes H5 and H7, and is the cause of epizootic among chicken with mortality reaching 100%. In 1999-2000 in the north-east of Italy in poultry farms spread low pathogenic avian influenza H7N1, which then transferred to the highly pathogenic and caused 413 outbreaks with deaths of more than 13 million birds of various species. In Australia, there were 5 outbreaks of highly pathogenic avian influenza, caused by a virus H7. During the last in 1997 in New South Wales from chickens were isolated two strains of influenza virus H7N4. Most strains from birds in the United States were isolated in specialized markets in the north-east of the country, and include mainly viruses of subtypes H7N2, H7N3, H5N2, etc. The vast majority of strains H7N2 belonged to the same line, first introduced in 1994, however, marked the introduction of new viruses and H7N2 and H7N3. Since 1997, the main line of H7N2 virus struck commercial livestock, at least three times, and has caused the most large-scale outbreak in Virginia in 2002. In the spring of 2003 in the Netherlands there was an epizootic of highly pathogenic influenza H7N7, which then spread to Belgium and Germany. In these countries, were devastated more than 33 million of chickens, as well as a significant number of sick and contacted with birds pigs. According to WHO, since 1959, all around the world were registered 21 epizootics of avian influenza, five of which were characterized by a significant coverage of farms and large economic losses. The paper analyzes the structural features of the surface proteins of viruses A/H7 of different origin and pathogenicity. The results of the study of epidemic variants of influenza A virus H7N9 that caused the outbreak in China in 2013 are described. The conclusion about importance of constant monitoring of the influenza A virus in bird populations is done.

Keywords: Influenza A virus, subtype H7, hemagglutinin, neuraminidase, bird, genome, pathogenic

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Author

page: 17-22 pp
DOI: 10.11134/btp.2.2013.3
К.Г. Ли
РГП «Республиканская коллекция микроорганизмов» КН МОН РК, г. Астана
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Abstract

Cellulose from biowaste is the most attractive substrate for the production of high quality products (e.g., fuel or plastics) by fermentation. However, the traditional processing of biomass is economically inefficient multi-step process. So far, no microorganisms able to perform single-stage fermentation (consolidated bioprocessing; CBP). The main technical problem in a cost-effective production of cellulosic biofuel is a need to reduce the cost of plant cell walls degrading enzymes (PCDE), which are necessary for the production of sugars from biomass. Several competitive low-cost technologies were developed for the production of PCDE in various host organisms such as E. coli, Zymomonas mobilis and some plants. There is possible heterologous expression in recombinant E. coli PCDE or Z. mobilis and successful consolidated bioprocessing in these microorganisms. In planta expression enables to simplify the enzyme production process and biomass processing leading to self-destruction of the plants cell walls. Although the future of the currently available technologies is difficult to predict their full implementation is likely through the integration of existing approaches to the development of breakthrough technologies.

Keywords: cellulases, heterologous expression, consolidated bioprocessing, biofuel.

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Author

page: 23-29 pp
DOI: 10.11134/btp.2.2013.4
Kuvat Momynaliev1, Vera Chelysheva2, Oksana Selezneva2, Andrey Larin2, Tatyana Akopian2, Dmitry Alexeev2, Veronique Le Berre3, Serguei Sokol3, Jean-Marie Francois3, Vadim Govorun2
1National Center for Biotechnology, Sh. Valikhanov str. 13/1, Astana, Kazakhstan
2Research Institute for Physico-Chemical Medicine, Malaya Pirogovskaya 1A, Moscow, Russia
3Biochips platform of Genopole, University of Toulouse, INSA, UPS, INP & INRA, 135, Avenue de Rangeuil, F-31077, Toulouse

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Abstract

Subinhibitory concentrations (sub - MICs) of antibiotics do not kill bacteria, but they are able to interfere with important aspects of bacterial cell function, such as adhesion to host cells, surface bacterial energy, susceptibility to host defense mechanisms, inhibition of enzyme function and toxin production. In order to understand how H. pylori copes with environmental stress and what facilitates the emergence of RIF mutants in H. pylori, we used DNA microarrays to compare the gene expression profiles of H. pylori in the presence and absence of subinhibitory concentrations of rifampicin (1/16 MIC (0.1 mg/L), 1/8 MIC (0,2mg/L), ¼ MIC (0,4 mg/L), and ½ MIC (0,8 mg/L). We found that еhe expression of 57 genes (of the 1,576 genes analyzed) was increased more than ≥ 1,5 – fold, and the expression of only 29 genes was decreased more than ≤ 1,5 –fold in significant way (p-value < 0,05), when H. pylori was treated with sub – MICs of RIF. No correlation was found between the sub - MICs of RIF and gene expression. We conclude that the alteration in the transcriptional pattern of H. pylori after the exposure to sub – MICs of RIF is mainly due to a direct interaction between rifampicin and the RNA polymerase β-subunit. Finally, we propose that subinhibitory concentrations of rifampicin may lead to an increase in the number of hypermutable cells in the H. pylori population.

Keywords: Helicobacter pylori, selection, microarray, rpoB.

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Author

page: 30-34 pp
DOI: 10.11134/btp.2.2013.5
П.В. Тарлыков1,2, М. Шоаиб3, А.Т. Кулыясов2, Е.М. Раманкулов2,4, В.В. Огрызько3
1Евразийский национальный университет им. Л.Н. Гумилева, г. Астана, Казахстан
2Национальный центр биотехнологии, г. Астана, Казахстан
3CNRS UMR 8126, Institut de Cancerologie Gustave Roussy, Villejuif, France
4Назарбаев Университет, г. Астана, Казахстан

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Abstract

Different combinations of histone posttranslational modifications recruit different interaction partners, such as chromatin regulators. Development of new methodologies that allow one to study protein-protein proximity in vivo may provide important information about protein dynamics and help to decipher remarkable complexity of histone crosstalk.

The new method is called PUB-NChIP (Proximity Utilizing Biotinylation with Native Chromatin ImmunoPrecipitation). PUB-NChIP is “in vivo biotinylation” approach to study chromatin in proximity to a protein of interest in which a nuclear protein of interest is fused to biotin ligase and co-expressed with a histone tagged with biotin acceptor peptide. In a model experiment biotin acceptor peptid was fused to specific histones and the ligase to Rad18, an E3 ubiquitin protein ligase associated with DNA repair. Biotinylation of specific histones in proximity to the Rad18 protein was observed. Biotinylation was used to isolate DNA associated with the subpopulation of histones proximate to the protein of interest. This preserves the ability to analyze post-translational modifications on the histones. Other advantages of PUB-NChIP include the ability to utilize histone variants associated with specific functional states (e.g., active or repressed chromatin), and the possibility to perform pulse-chase experiments to monitor chromatin fate after it was in proximity with the nuclear protein of interest.

The application of the new methodology to study histone posttranslational modifications, developed in our work, may shed a new light on cellular mechanisms of chromatin remodeling, and have a general relevance for our understanding of the mechanisms of base excision repair and epigenetic reprogramming.

Keywords: chromatin, epigenetics, biotinylation, western-blot, mass spectrometry

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Author

page: 35-41 pp
DOI: 10.11134/btp.2.2013.6
К.П. Аубакирова, М.Е. Омашева, Н.А. Рябушкина, Л.В. Береснева, Н.Н. Галиакпаров
РГП «Институт биологии и биотехнологии растений», Министерство образования и науки Республики Казахстан, Комитет науки, г. Алматы
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Abstract

Programs and Regulations of Kazakhstan Government are aimed to recover the vineyards area from 9500 to 26 000 ha, productivity up to 220 000 tons. The success of viticulture industry in Kazakhstan depends on selection and introduction of modern commercial grapevine cultivars resistant to biotic and abiotic stresses of the agro-climatic zones of Kazakhstan. Kazakhstan breeders developed traditionally a number of table and wine V. vinifera varieties, some of which declared as cold resistant and resistant to the most destructive grapevine pathogens. Modern characteristics of grape varieties, along with morphological and agronomic features include molecular genetic profiles based on molecular markers. Microsatellites are most widely used for the identification and differentiation of grapevine varieties. Researchers have identified a minimal set of microsatellites, allowing optimal differentiation of varieties in germplasm collections.

In this paper, for the first time for grape genotyping the method with loci markers VVS2, VVMD5, VVMD7, VVMD27, ssrVrZAG62 and ssrVrZAG79 was combined and optimized using three universal unspecific oligonucleotides with different fluorescent dyes. This technique has been used for genotyping a number of Kazakhstan’ varieties, their parents and some European reference varieties. For each of the 6 loci the conversion factor has been calculated to determine the exact size of the alleles. As a result, the sizes of alleles determined in the experiment were adequate to the alleles in the European database Swiss Vitis Microsatellite Database. The analysis confirmed for Kazakhstan’s varieties parent-offspring relationship. The method will be used for genetic certification of grapevine varieties in Kazakhstan.

Keywords: grape, genotyping, ssr markers, DNA, PCR.

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Author

page: 42-46 pp
DOI: 10.11134/btp.2.2013.7
Д.С. Тагиманова, А.Ж. Ергалиева, О.Б. Райзер, О.Н. Хапилина
РГП «Национальный центр биотехнологии» КН МОН РК, Казахстан, г. Астана
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Abstract

Drought - one of the main reasons why the productivity of plants. To ensure that agriculture from losses during drought years, you need to have resistant to moisture deficit varieties of wheat, barley and other crops. Plants under drought meant their ability to most efficiently use the water at high temperature, low humidity, soil and air, and in these conditions give high yield and good quality products. In terms of agronomic drought - is not the survival of plants under drought conditions, and the ability to maintain a relatively high level of productivity in the shortage of water [1]. Throughout the world, there are many publications devoted to the problem of obtaining varieties of spring wheat that are resistant to drought. The studies assessed the breeding material for drought tolerance. To assess the sustainability of breeding material for drought used a modified test procedure wheat in vitro. Used as explants mature embryos of wheat seeds. An indicator of the stability of the samples in this experiment was the manifestation of shoot in harsh selective conditions. Characterized studied variety of medium and high resistance to selective conditions in vitro. Laboratory methods are the most simple methods of mass estimation of drought. An indicator of the stability of genotypes to osmotic stress is a degree of depression in the accumulation of dry matter sprouts with increased osmotic pressure. The less inhibited growth and biomass accumulation wheat seedlings in a sucrose solution as compared with the control, the more stable the sample.

Keywords: spring wheat, variety, in vitro, drought tolerance, testing

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Author

page: 47-52 pp
DOI: 10.11134/btp.2.2013.8
К.Н. Мукантаев, А.В. Шустов, Ы. Сыдыкнаби, Ш. Байдосова, К.К. Муканов
РГП «Национальный центр биотехнологии» КН МОН РК, г. Астана
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Abstract

An important problem for prevention of the bovine leukemia is an effective serological diagnostics. The main methods of serologic diagnostics utilized in the programs for the prevention and eradication of the disease are the immune diffusion reaction and enzyme-linked immunosorbent assay.

For diagnostic test kits used in the detection of bovine leukemia in cattle traditional antigen is an inactivated virus grown in the FLK cell cultures. This antigen often demonstrates cross-reactivity with antibodies to other animal viruses. One considered reason for the decrease in specificity is possible coinfection of the FLK cell cultures with virural contaminants present in the calf serum used in the culture medium. To increase the specificity of diagnostic tests a method of producing a recombinant antigen gp51 was developed using baculovirus expression system. However, this technology does not completely solve the existing problems, since it involves the use of expensive culture media and does not eliminate complex product purification. Methods of genetic engineering have already provided a variety of recombinant antigens for the use in diagnosis of veterinary infections. Recombinant DNA technology allows to obtain highly purified stable formulations of recombinant glycoprotein gp51 of the bovine leukemia virus.

The aim of this work is to develop a bacterial expression system for the gp51 protein of the bovine leukemia virus and to study the gp51 diagnostic properties in ELISA.

We designed a synthetic gp51 gene and synthesized its nucleotide sequence, 858 bp in length. The resulting gene was cloned into the construct for the recombinant protein expression pET32/gp51. The E.coli strain BL21(DE3) was transformed with the pET32/gp51 and the method for purification of the gp51 antigen was developed. Culture conditions and induction parameters have been optimized to increase the yield of the recombinant gp51. A method of ELISA was developed using the recombinant gp51. The ELISA results were fully congruent to the results of the routinely used diagnostic test for bovine leukosis – immune diffusion precipitation reaction (RDP).

Keywords: virus, leukemia, gp51, recombinant antigen, ELISA, immune diffusion precipitation.

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Author

page: 53-55 pp
DOI: 10.11134/btp.2.2013.9
А.М. Жусупова, С.К. Барбасова, О.А. Тен
Филиал РГП «Национальный центр биотехнологии» КН МОН РК, г. Степногорск
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Abstract

L-лизин является незаменимой аминокислотой, необходимой для питания животных и человека. В практике животноводства целесообразно применение чистого лизина промышленного производства. Перспективным является получение лизина путем микробиологического синтеза. Был проведен подбор питательной среды с оптимальными концентрациями составляющих компонентов, обеспечивающих максимальное накопление лизина при культивировании штамма-продуцента Brevibacterium sp. шт. 92. Оценку качества оптимизации проводили методами биохимического анализа культуральной жидкости: определение концентрации сухой биомассы фильтрационным методом, определения редуцирующих веществ - методом Бертрана. Определение массовой концентрации аминокислот проводили методом высокоэффективной жидкостной хроматографии (ВЭЖХ). Выбрана оптимальная среда ФЛ следующего состава, г/л: сахароза - 100,0; KH2PO4 - 5,0; MgSO4х7H2O - 5,0; биотин - 1,3; тиамин - 3,3; дрожжевой автолизат - 10,0 мл/л, треонин - 6,6. Концентрация лизина в культуральной жидкости Brevibacterium sp. шт. 92 на среде ФЛ составила 98,7 мг/г. Финансирование работы осуществлено в рамках проекта «Разработка технологии производства кормовой добавки на основе использования лизина, треонина и метионина» по межгосударственной целевой программе Евразийского экономического сообщества «Инновационные биотехнологии» на 2012-2014 гг.

Ключевые слова: лизин, штамм-продуцент, высокоэффективная жидкостная хроматография.

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Author

page: 56-58 pp
DOI: 10.11134/btp.2.2013.10
А.Н. Ермолаева, У.Ж. Алгожина, О.А. Тен, Д.С. Балпанов
Филиал РГП «Национальный центр биотехнологии Республики Казахстан», МОН РК,г. Степногорск
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Abstract

The data on the isolation and study of the biochemical properties of probiotic cultures kind of Bifidobacterium bifidum. As materials for the isolation of probiotic cultures baby feces samples used first days of life. Fecal samples were obtained during the stay of children up to 7 days of life in the hospital. We selected 10 samples of feces. From the gastrointestinal tract baby seven days of life have been allocated four individual isolates. We studied the cultural-morphological, physiological and biochemical properties of the isolates. When the microscope met the still curved, club-shaped rods arranged singly or gathered in the chain. The dispute did not form. Cells were 0,5 micron. Anaerobes. Acid-fast. The optimum growth temperature is 37-41ºC. Grew well on the medium Bifidobacterium broth (bouillon for bifidobacteria). Did not grow at pH 4,5. Catalase. Gram. Based on the data from the study of physiological and biochemical properties isolates active assimilate a wide variety of carbohydrates: galactose, glucose, lactose, mannose, sucrose, fructose, to form a free acid gas catalase did not form. Do not consume arabinose, xylose, rhamnose, ribose, salicin and cellobiose.

Keywords: Bifidobacterium bifidum, kind, isolates, штамм, feces samples, carbohydrates

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