Construction of stably maintained non-mobilizable derivatives of RSF1010 lacking all known elements essential for mobilization

<p>Abstract</p> <p>Background</p> <p>RSF1010 is a well-studied broad-host-range plasmid able to be mobilized to different bacteria and plants. RSF1010-derived plasmid vectors are widely used in both basic research and industrial applications. In the latter case, exploiting of mobilizable plasmids or even the plasmids possessing negligible mobilization frequency, but containing DNA fragments that could promote conjugal transfer, is undesirable because of biosafety considerations. Previously, several mutations significantly decreasing efficiency of RSF1010 mobilization have been selected. Nevertheless, construction of the RSF1010 derivative lacking all known loci involved in the conjugal transfer has not been reported yet.</p> <p>Results</p> <p>Novel non-mobilizable derivatives of RSF1010 lacking all known DNA sequences involved in the mobilization process have been obtained due to the exploiting of λRed-driven recombination between the plasmid and a constructed <it>in vitro </it>linear DNA fragment. To provide auto-regulated transcription of the essential replication gene, <it>repB</it>, the plasmid loci <it>oriT</it>, <it>mobC </it>and <it>mobA </it>were substituted by the DNA fragment containing P_{<it>lac</it>UV5}→<it>lacI</it>. Mobilization of the obtained RSFmob plasmid was not detected in standard tests. The derivative of RSFmob with increased copy number has been obtained after <it>lacI </it>elimination. High stability of both constructed plasmids has been demonstrated in <it>Escherichia coli </it>and <it>Pantoea ananatis</it>. Design of RSFmob allows easy substitution of P_{<it>lac</it>UV5}by any desirable promoter for construction of novel derivatives with changed copy number or host range.</p> <p>Conclusion</p> <p>Novel non-mobilizable derivatives of RSF1010 lacking all known DNA sequences involved in the mobilization process and stably maintained at least in <it>E. coli </it>and <it>P. ananatis </it>have been constructed. The obtained plasmids became the progenitors of new cloning vectors answering all biosafety requirements of genetically modified organisms used in scale-up production.</p

Синтез ацильных производных тиазоло[5,4-b]индола.

2-Amino-4-acetylthiazolo[5,4-b]indole and its bromosubstituted analogue contain a free amino group and are initial compounds for chemical modification in order to obtain on their basis functional derivatives having high therapeutic activity. By the interaction of these 2-aminо-4-acetylthiazolo[5,4-b]indoles with succinic and maleic anhydrides, the corresponding imides and amides were obtained, which are of interest for use in order to create on their basis antihypoxic and actoprotective agents.2-амино-4-ацетилтиазоло[5,4-b]индол и его 7-бромзамещенный аналог содержат свободную аминогруппу и являются исходными соединениями для химической модификации с целью получения на их основе ряд функциональных производных, обладающих терапевтической активностью. Взаимодействием указанных 2-аминотиазоло[5,4-b]индолов с янтарным и малеиновым ангидридами были получены соответствующие имидои амидопроизводные, представляющие интерес для создания на их основе лекарственных средств антигипоксического и актопротекторного действия

Синтез S-аминоалкилпроизводных 1,2,4-триазино[5,6-b]индола

Condensed indole derivatives of the 1,2,4-triazino[5,6-b]indole group show high biological activity and are of interest for the development of pharmaceuticals that have antihypoxic and antisurditant effects. By S-aminoalkylation reactions on a thione group, S-aminoalkyl derivatives of 1,2,4-triazino[5,6-b]indole-3-thione that were subject to hydrobromination in order to obtain the form of hydrobromides were synthesized, isolated and characterized.Конденсированные производные индола группы 1,2,4-триазино[5,6-b]индола обладают высокой биологической активностью и представляют интерес для создания лекарственных средств, проявляющих антигипоксическое и антисурдитантное действия. При проведении по тионовой группе реакции S-аминоалкилирования синтезированы, выделены и охарактеризованы S-аминоалкилпроизводные 1,2,4-триазино[5,6-b]индола, которые подвергали гидробромированию для получения их в форме гидробромидов

THE GROUP COMPOSITION OF HUMUS AND FRACTIONS OF FINE-DISPERSED PHASE IN THE SODDY-PODZOLIC MEDIUM-LOAMY SOIL IN A LONG-TERM EXPERIMENT

The group composition of humus in microaggregates (colloidal, pre-colloidal, fine, medium and coarse silt) of the soddy-podzolic medium-loamy soil has been first studied in three five-course grain-tilled crop rotations in the long-term experiment of V.V. Dokuchaev Soil Science Institute. It is shown that in the trial with a higher dose of fertilizers (N150P150K180 kg/ha/year + 120 t/ha of manure) the humus content increases in soil after the first rotation as compared to that with average fertilizer doses (N60P60K60). This is explained by the content of carbon bond to microaggregates of medium and coarse silt. At the same time both experimental variants reveal deterioration of the microstructure including the decrease in the content of medium silt, accumulation of pre-colloidal and fine-dispersed fractions. It becomes evident that the best microstructure is observed in the variant enriched with fertilizer: it contains a great amount of silty fractions and humus components in the form of humates and nonhydrolyzed residues. Moreover, both experimental variants show a tendency towards decreasing the content of humates bond to silty fractions. The intensive system of fertilization in three rotations allowed maintaining the stable content of humus in the fraction of medium silt, the latter being disturbed to a lesser extent than that in the variant without manure application

The monetization coefficient of the region and its impact on the development of a territorial unit’s economy

В последние годы в мировой и отечественной экономической литературе широко обсуждается вопрос о роли финансовой системы в развитии национальных экономик. Статья посвящена анализу влияния финансовой глубиныэкономики на уровень ее развития в отдельных регионах Российской Федерации. Отметим, что в научном обороте понятие «финансовая глубина экономики» является еще недостаточно изученным, и, как следствие, анализу влияния отдельных индикаторов «финансовой глубины» на уровень развития территориальных образований внутри государства уделяется недостаточно внимания. Методологической базой исследования явились теоретические положения макроэкономической теории, описывающие наличие взаимных связей между уровнем развития финансового сектора и экономическим ростом. В качестве цели настоящего исследования определено изучение возможного влияния одного из индикаторов финансовой глубины региональной экономики (коэффициента монетизации замкнутых территориальных образований внутри государства, имеющего федеративное устройство) на уровень развития экономики соответствующего региона. Информационную базу исследования составляют данные по 20 субъектам РФ с наибольшей и наименьшей величиной статистического показателя «средства организаций, банковские депозиты (вклады) и другие привлеченные средства юридических и физических лиц». Результаты анализа свидетельствуют: финансовая глубина экономики региона по индикатору «коэффициент монетизации» не превышает 50%, за исключением двух ключевых субъектов (Москвы и Санкт-Петербурга), в то время как в странах с развитой экономикой этот показатель достигает уровня 200%. Рассчитанный коэффициент детерминации указывает на несущественное влияние фактора (монетизация конкретного субъекта РФ) на вариацию результативного показателя (ВРП в расчете на одного жителя субъекта выборки). Таким образом, воздействие одного из индикаторов финансовой глубины региональной экономики – коэффициента монетизации замкнутых территориальных образований – на уровень развития экономики соответствующего региона не находит в российской практике подтверждения.In recent years, the role of the financial system in the development of national economies has been widely discussed in the global and national economic literature. This article analyzes the influence the financial depth of economy has on the level of its development in some Russian regions. Notably, the notion of «financial depth of economy» is not yet sufficiently studied, and therefore, the analysis of the impact, which individual indices of «financial depth» have on the level of development of territorial entities within the state, has not received sufficient attention. The methodological basis of the study includestheoretical foundations of macroeconomic theory, describing the existence of mutual relations between the level of development of the financial sector and the economic growth. This study aims to study the possible impact of one of the indices of the financial depth of the regional economy – the coefficient of monetization of closed territorial formations within the state, which has a federal structure – on the level of the region’s economic development. The information base of the research consists of the data on 20 constituent entities of the Russian Federation with the highest and lowest value of the statistical index «funds of organizations, bank deposits (deposits) and other borrowed funds of legal entities and individuals». The results show that the financial depth of the region’s economy according to the index «monetization coefficient» does not exceed 50%, except for two key subjects (Moscow, St. Petersburg), while in the countries with developed economies this index reaches the level of 200%. The calculated determination coefficient indicates an insignificant impact of the factor (monetization of a specific RF entity) on the variation of the effective index (GRP per capita of the sample subject). Thus, the impact of one of the indices of the financial depth of the regional economy – the coefficient of monetization of closed territorial formations – on the level of economic development of the region does not find a corresponding confirmation in the Russian experience

Construction of stably maintained non-mobilizable derivatives of RSF1010 lacking all known elements essential for mobilization-1

<p><b>Copyright information:</b></p><p>Taken from "Construction of stably maintained non-mobilizable derivatives of RSF1010 lacking all known elements essential for mobilization"</p><p>http://www.biomedcentral.com/1472-6750/7/80</p><p>BMC Biotechnology 2007;7():80-80.</p><p>Published online 21 Nov 2007</p><p>PMCID:PMC2200642.</p><p></p>e were analyzed. 1, 2, 3 – 0.5, 2.0 and 5.0 μl of the DNA probe isolated from clone 1 of MG1655/RSF1010; 4, 5 – 2.0 μl of the DNA probes isolated from clones 2 and 3 of MG1655/RSF1010; 6, 7, 8 – 2.0 μl of the DNA probes isolated from 3 independent clones of MG1655/RSFmob; 9, 10, 11 – 2.0 μl of the DNA probes isolated from 3 independent clones of MG1655/RSFmob-I. Relative copy number was calculated as the ratio of the DNA amounts in the upper bands (5.9 kb EcoRV-EcoRV fragment). Electrophoretic patterns for stationary phase are represented as an example