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21.
Hydrogeological and speleological research of the spring cave Slatinski Izvor and its recharge are (Republic of Macedonia)
Biljana Gichevski, 2016, doktorska disertacija

Opis: This thesis represents a comprehensive study, giving first extensive information on the hydrogeological and speleological characteristics of the spring cave Slatinski Izvor and its catchment area. The researched spring is located in the Poreče basin of West-Central Macedonia, in the river valley of Slatinska Reka. The wider area around the spring belongs to two tectonic units. The Slatinski Izvor spring is situated on the Pelagonian horst anticlinorium, which is composed of carbonate rocks. The second is the Western Macedonian zone, and it is composed of non-carbonate rocks. The spring represents a significant potential for water supply for the settlement Slatina. Prior to this research only some basic information about the karst aquifer in Precambrian dolomite marbles was available. Therefore, my goal was to ensure better understanding of the functioning of the karst system in the recharge area of the spring, assessment of its vulnerability to various pollution sources, as well as study of karst development of the area. Because karst aquifers have unique hydrogeological characteristics and specific nature, they also require specifically adapted investigation methods. In order to reach the main goal, speleological investigations, hydrological methods, hydrochemical methods and tracer test were used. A monitoring network was combined with regular samplings for major and trace elements analysis, which enabled considerable advances in understanding the functioning of the karst system. The data from two-year period (December 2011 – November 2013) was compared and analyzed. The controlling of the karst development in the study area mostly depends on the tectonic and geologic conditions. Incision of Slatinska Reka is the main controlling factor in cave development by lowering the base level of karst terrains. All investigated caves (Gorna Slatinska, Slatinska II, Ovčarska Peštera, Puralo, Slatinski Izvor) have “normal” epigenic karst development. Slatinski Izvor is the youngest cave. The Slatinski Izvor spring serves as a cave entrance for the same cave and, represents the outflow of groundwater from the karst system. A conceptual model of the karst system of the study area was developed. The Slatinski Izvor spring has typical karst hydrological regime. The karst system is well developed by conduits and rains infiltrated into vadose zone flow rapidly towards the spring. The travel time of low mineralized water within the system corresponds well with the results from an artificial tracer test. Performed artificial tracer test confirmed that the Slatinski Izvor spring is recharged by allogenic stream. The dominant apparent flow velocity of 250 m/h, a single peak of the tracer breakthrough curve and more than 87% of tracer recovered indicate a rapid conduit flow and high vulnerability of the observed drinking water source. Analysis of spatial and temporal variations of physical and chemical parameters show that waters in the study area had different origin and were transmitted along different flow paths. The origin of the water of the Slatinski Izvor spring is from non-carbonate area, but the influence of mixed limestone-dolomite sequence in the carbonate area is significant. The results point out to short residence time of the water in the karst aquifer. The anthropogenic impact in the study area is insignificant which is reflected in the good water quality. In order to preserve it, land surface zoning was performed in terms of groundwater and spring water vulnerability to pollution according to the hydrogeological research. A comprehensive research with a combined use of various research tools presents an innovative approach and a new contribution to the Macedonian karstological science. Applied methods proved to be successful for to study of the karst system. Finally, the results of the study have clear applicative significance in terms of drinking water management.
Ključne besede: karst hydrology, cave, speleological investigation, hydrochemistry, tracer test, spring Slatinski Izvor
Objavljeno: 14.10.2016; Ogledov: 1463; Prenosov: 24
.pdf Polno besedilo (11,01 MB)

22.
Transformacija antičnih motivov v slovenski književnosti devetnajstega in dvajsetega stoletja
Urša Prša, doktorska disertacija

Opis: Antična mitologija že stoletja navdihuje evropski prostor ter se v njem na različne načine transformira in interpretira v različnih vrstah umetnosti. Antični motivi so odmevali v slovenski književnosti od srednjega veka pa vse do sodobnosti. Slovenska literatura jih je v različnih fazah svojega razvoja sprejemala ali zavračala ter iz njih oblikovala svoje motivno-tematske zasnove. Posebna pozornost te doktorske disertacije je posvečena tipom literarne recepcije, ki se je udejanjila v slovenskem slovstvu romantike in v dvajsetem stoletju, ko so antični liki in motivi postali simboli in prispodobe moderne subjektivnosti. Po krajšem pregledu tuje in domače znanosti o mitologiji s poudarkom na teoretski obravnavi mita v literaturi sledi opazovanje recepcije antične motivike v slovenskem in evropskem literarnem prostoru od antike naprej. Posebna pozornost bo namenjena motivom Medeje, Odiseja in Orfeja v izbranih delih slovenske književnosti 19. in 20. stoletja, saj je vsak lik s svojo univerzalnostjo in enkratnostjo pomembno vplival na slovensko literaturo. Ob pomoči medbesedilne metode so analizirani namen avtorjeve uporabe izbranega antičnega motiva, njegova vloga, interpretacija in morebitna transformacija ali aktualizacija.
Ključne besede: antični motivi, transformacija, slovenska književnost, Medeja, Odisej, Orfej
Objavljeno: 11.10.2016; Ogledov: 1442; Prenosov: 163
.pdf Polno besedilo (2,52 MB)

23.
THE AUTOBIOGRAPHICAL EXPERIENCE OF MIGRATION IN THE LITERARY WORKS OF WOMEN WRITERS OF THE SLOVENIAN LITERARY POLYSYSTEM
Megi Rožič, 2016, doktorska disertacija

Opis: People have travelled and migrated since the early periods of history and the phenomenon of migration, defined as “the movement of large numbers of people, birds or animals from one place to another” (Oxford Advanced Dictionary, 7th ed.) has a history of hundreds of years (Pourjafari, Vahidpour, 2014) and is by no means new. But the way of travelling, migrating and mobility in contemporary reality does have many specific aspects and recently it has been studied in new ways, with new concerns. The experience of migration in the contemporary world is a fundamental characteristic of human societies. “It is a system in which the circulation of people, sources and information follows multiple paths. The energy and barriers that alter the course or deflect the contemporary patterns of movement have both obvious and hidden features. While nothing is utterly random, the consequences of change are often far from predictable” (Papastergiadis, 2000: 1). This unpredictability and multidirectionality has led to changes in approaches to the study of migration in recent decades. In the last few decades the study of migration has increasingly been accompanied by a tendency to study it on the individual level − at the level of personal life stories (Milharčič Hladnik, 2007). These are also markedly expressed in the medium of literature. The present dissertation presents literary oeuvres of seven women writers, who thematise the autobiographical experience of migration, it offers an individually, woman-centred experience and view of migration. In selected oeuvres personal views are expressed on that experience, different strategies of coping with life in new realities and regarding relationships in these environments. The thematisation of the experience of migration in selected oeuvres is also connected with a problematisation of other concepts: the concepts of belonging, borders, nation-state, culture and language. It is also vitally connected with the personal identity construction of the lyrical subjects and literary characters and also because of the autobiographical character of selected literary works of the literary artists themselves. The experience of migration in selected literary works leads to unique identity formations, which in themselves combine elements of different cultural backgrounds and traditions. In their literary works these selected women writers also shape their relationship toward time and space dimensions, tradition and interpersonal relationships through the experience of migration. The experience of migration, the relocation of the subject and a change of the geographical area in selected oeuvres, does not only represent a change of the geographical position: it also allows a deviation from other rigid and seemingly fixed and unvarying patterns and virtual realities that accompany human life. Migration can also present an alienation effect from traditionalisms and determinants that define human lives. In the literary oeuvres of these selected women writers, migration is only in part tied to the traditional concepts related to migrants, with the loss of roots and rupture with the place of origin. In the selected literary oeuvres, migrants are rarely considered to be uprooted and unable to find their anchor or confidently start a new chapter in their lives in a new environment. Migration is mostly connected with the possibility of expanding the horizon of insights and perspectives of looking at life, with the acceptance of its complexities, ambiguities and incompleteness. Mostly, the experience of migration is seen as a new, creative option, which opens and examines the wide range of other issues and dilemmas. The condition of uprooted loss has traditionally negative connotations, but in the selected oeuvres vagueness and fluidity allow a real insight into the real, complex nature of life and human existence.
Ključne besede: The experience of migration, women writers, Maruša Krese, Ifigenija Zagoričnik Simonović, Brina Švigelj-Mérat − Brina Svit, Gabriela Babnik, Stanislava Chrobáková Repar, Erica Johnson Debeljak, Lidija Dimkovska, literary polysystem, nomadic entity, locational feminism, fluid identity, transnationalism, transculturalism
Objavljeno: 11.10.2016; Ogledov: 1800; Prenosov: 246
.pdf Polno besedilo (1,78 MB)

24.
General framework for the conservation of historical rural landscape. Case study of Qadisha Valley in Lebanon.
Bachir Zarif Keyrouz, 2016, doktorska disertacija

Opis: The international community has reacted to the challenges faced by historic cities and cultural landscapes. In 2011, UNESCO adopted the international recommendation on the Historic Urban Landscape in order to draw the attention of governments and local communities to the need to improve the management of historic urban areas. The aim of the present dissertation is to expand the UNESCO definition to Historic Rural Landscapes, and to explore how the proposed methodology could be applied to the management of rural areas. The dissertation takes the Qadisha Valley in Lebanon as an example, where some elements have already been recognised for their outstanding universal value. It is considered however that these historic-cultural elements are part of the larger context of the Qadisha Valley, which has its specificity and its qualities that merit being recognised. It is also considered that the management of the elements already recognised by UNESCO can only be successfully safeguarded if the management is understood in the context of the entire valley. Consequently, the thesis is articulated taking into account the following parameters: 1. UNESCO has already set the restoration plan to solve the problems of Qadisha Valley. Based on this plan, the question can be raised whether or not the rural planning in Qadisha is well managed at the present? 2. In case Qadisha Zone is not well conserved, what improvements could be brought about and implemented in order to safeguard the cultural heritage and historical rural landscape, and what should be the guiding policies of restoration? 3. Once a restoration plan is implemented in Qadisha Zone, what management plan should be brought up and implemented to safeguard its cultural heritage and historical rural landscape? 4. How can conservation of cultural heritage in Qadisha Zone be done culturally and in a socially acceptable way? 5. What sectors of the nation are to be involved in the Management plan? How can the MP preserve and restore Qadisha Zone? The aim of the dissertation has been to examine the possibilities of involvement of the local community and relevant authorities in a more structured management of their properties within the context of the entire valley and its exceptional historic and landscape qualities. It is hoped that the study can generate more interest in the management of the rural context within the increasingly globalising society, and draw attention to the qualities of historic rural landscapes as the setting for an improved quality of life.
Ključne besede: Historic Rural Landscape, Qadisha World Heritage, Heritage Community, Sustainable Tourism, Strategic Urban Management, Outstanding Universal Values & Conservation Process.
Objavljeno: 07.10.2016; Ogledov: 1461; Prenosov: 27
.pdf Polno besedilo (58,03 MB)

25.
Building the shared dimension of the rural landscape: tools, principles and methods
Caterina Groli, 2016, doktorska disertacija

Opis: This research investigates the rural landscape of the lagoon of Venice and its surrounding territory focusing on the forms of exploitation of its natural resources, the transformations that occurred throughout the course of history and the consequent formation of its primary economical sector. New economic sectors, new agricultural policies and other spatial and sectorial drivers transformed the primary sector itself which changed and adapted its productive schemes to the main economic patterns, heavily affecting the former rural asset of the landscape of the lagoon by changing its demographic, economic activities and land use practices. The study carries out a systematic and detailed analysis of all economic, cultural and social features that have interacted and impacted this specific landscape, relying mainly on the comparison of historical and current land-use maps through three different historical spans: 1823-1840 (pre-industrialization phase), 1933-1966 (industrializing process, considered as a determining factor of the transformation of this landscape) and 2013 (present post-industrialized condition). The maps, combined with relative data, are compared with the aid of the Geographical Information System (GIS) computer software. As a result, a set of new maps has been obtained, which point out the historical landscape permanencies, defined through in situ surveys on the territory. A multitude of other archival documents from direct and indirect sources have been examined in order to build up a sound and detailed socio-economic context and the overall historical background. Resources, goods and services provided by the rural landscape and its development are an important part of regional and EU policy instruments and also an essential factor of identity and growth for local communities. The consumption of resources, their availability and maintenance over time and accessibility through property regulations are all factors that shape the common and public dimension of the rural landscape, which represents another study challenge of this work. This study proposes the establishment of the rural park of the lagoon of Venice, through the implementation of the multi-functional role of agriculture, for the fulfilment of two fundamental conditions of the peri-urban rurality: the need of safeguarding the landscape and its significance, by preserving the authenticity and integrity of the rural characters and the dynamism of the primary sector, which is strictly related to the survival of the rural landscape.
Ključne besede: Rural landscape of the lagoon of Venice, UNESCO Management Plan 2012–2018, European Landscape Convention, Common Agricultural Policy (CAP), Italian Constitution, Cultural Heritage and Landscape Code, authenticity and integrity of the rural heritage, intangible heritage, landscape evaluation, land-use cartography, Geographical Information System (GIS), fish-farming, multifunctional agriculture, new rural community, common pool resources, public goods, club goods, property rights, rural park
Objavljeno: 04.10.2016; Ogledov: 1074; Prenosov: 22
.pdf Polno besedilo (21,33 MB)

26.
SYMBIOSIS ECOLOGY OF SELECTED SCYPHOZOA
Lucija Raspor Dall'Olio, 2016, doktorska disertacija

Opis: Scyphozoa with symbionts have an advantage in oligotrophic environments due to the additional source of nutrients provided by their symbiontic algae, just as corals have benefits from their symbionts. The literature, however, has thus far devoted far less attention to the association between scyphozoan hosts and Symbiodinium sp. than it does to corals. This thesis investigated the identity of symbionts from scyphozoan medusae (Cotylorhiza tuberculata, Phyllorhiza punctata and Cassiopea xamachana) using a phylogenetic approach. Two scyphozoan species, P. punctata and C. tuberculata, were sampled over the Mediterranean Sea, while Cassiopea xamachana was sampled in the Atlantic Ocean. Symbionts were identified from live medusae and their identity and phylogenetic relationships were determined by analysing two nuclear markers, ITS2 and 28S rDNA, from symbionts. Symbiodinium sp. sequences belong to clades A, B, and C based on markers ITS2 and 28S rDNA. Moreover, individual medusae host only one type Symbiodinium (A, B or C). Host species from the Mediterranean Sea hosted Symbiodinium from clade A and B (C. tuberculata) or only from clade A (Phyllorhiza punctata), while the host from the Atlantic Ocean (C. xamachana) hosted Symbiodinium from clade B or C. The phylogeography of C. tuberculata medusae was analysed using mtCO1. All the haplotypes sampled over the Mediterranean Sea were clustered together without any sign of phylogeographic structuring.
Ključne besede: Scyphozoa, Cotylorhiza tuberculata, Symbiodinium sp., symbiosis, Mediterranean Sea, phylogeography, CO1, 28S rDNA, ITS regions
Objavljeno: 03.10.2016; Ogledov: 1397; Prenosov: 155
.pdf Polno besedilo (3,20 MB)
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27.
Intelektualci in institucije v slovenski zgodovini 17. stoletja
Andrej Koritnik, 2016, doktorska disertacija

Opis: Disertacija želi razkriti in sintetično povezati v kompleksno historiografsko sliko zgodbo 17. stoletja, ki jo razume kot integralni del novega veka, zato je vpogled v 16. stoletje, ko so delovali protestanti (kot posebna forma intelektualcev), in v 18. stoletje, ko so se stare institucije umikale novim in je razsvetljenska filozofija preplavila intelektualne kroge kakor tudi ljudsko kulturo, nujen, da lahko razgrnemo socialno, kulturno in intelektualno zgodovino na Slovenskem v 17. stoletju. Tembolj zato, ker za tedanji čas še ne moremo govoriti o konceptu »nacionalnega«, ki se je povezal z ljudskimi jeziki – pri nas seveda s slovenščino, temveč o počasnem, toda »revolucionarnem« spreminjanju starih družbenih in kulturnih vzorcev, v katerih sta védnost in intelekt legitimno izhodišče za historiografsko, a vseeno interdisciplinarno analizo in sintezo obravnavanega časa. V luči spora med Descartesom in Vicom ter njunima nasprotnima umevanjema zgodovine bo treba premisliti tudi, kako so ideje iz zgodovinopisja vplivale na splošno zgodovino idej, ki se je »dogajala« tudi med Alpami in Jadranom, ter kakšno težo in vpliv imajo »predrazsvetljenski« časi, o katerih pravzaprav ne vemo veliko, na našo sodobnost. Prikazali bomo njeno raznotero, zapleteno podobo kot preplet vseh mogočih silnic – v šolstvu, državni upravi, v fevdalnih razmerjih, v verskih sporih, jezikovni ideologiji, v gospodarstvu in drugod –, skozi katere sledimo liku intelektualca, bodisi kot člana neke institucije védnosti ali pa »osamelca«.
Ključne besede: Intelektualec, institucija, védnost, zgodovina védnosti, 17. stoletje, intelektualna zgodovina, kulturna zgodovina, literarna zgodovina, protestantizem, protireformacija, katoliška obnova, znanstvena revolucija, univerze, osnovne šole, akademije
Objavljeno: 14.09.2016; Ogledov: 1141; Prenosov: 108
.pdf Polno besedilo (1,74 MB)

28.
ECOLOGICAL STUDIES OF EPIKARST COMMUNITIES IN ALPINE AND PRE-ALPINE CAVES
Federica Papi, 2016, doktorska disertacija

Opis: The karst ecosystem shows heterogeneity and variability of geology, hydrology, morphology and ecology in space and time. Chemical composition of water in karst plays a crucial role not only in dissolution of karst rocks and deposition but also in ecological processes. The heterogeneous nature of karst aquifers leads to difficulties in predicting groundwater flow and contaminant transport direction and travel times. For its position at the top of karst, epikarst represents the interface between surface and underground. Epikarst ecology, biodiversity and fauna have rarely been systematically studied. The typical absence of enterable passages makes necessary an indirect approach. In this research, epikarst biodiversity in relation to environmental conditions was studied in seven Alpine and Pre-Alpine caves, located at different altitudes, in Slovenia and in Italy: Snežna jama na planini Arto (1556 m a.s.l.), Jama pod Babjim zobom (860 m a.s.l.), Zadlaška jama (300 m a.s.l.) and Pološka jama (500 m a.s.l.), in Slovenian Alps and Pre-alps, and Grotta A del Ponte di Veja (600 m a.s.l.), Covolo della Croce (875 m a.s.l.) and Grotta di Roverè Mille (1005 m a.s.l.) in Lessinian Massif. In these caves, percolating water was monthly monitored for fauna in the period of one year. Temperature, discharge, conductivity and pH were measured at the same time, and water samples for the laboratory analyses of total hardness, concentrations of various ions (calcium, chlorine, nitrate, sulphate and phosphate) and dissolved organic carbon were taken. 66 aquatic and terrestrial, mostly hypogean, species were found. Aquatic fauna was dominant in all caves, with the exception of Covolo della Croce. Caves located at higher elevations harbour less diverse communities. No animals were found when discharge was very low and in correspondence of moonmilk formations. The most abundant and diverse group was Copepoda, with thirteen different species and 776 individuals at different maturity stadia, totally representing 61% of the community (between two and three copepod species per cave in Slovenia and between one and six per cave in Italy). The 62% of copepod species were stygobionts. Some copepod species (e.g. Speocyclops infernus) have wide ecological tolerance to environmental variables, other have more restricted tolerances (e.g. Lessinocamptus insoletus). With regard of copepods, there was a good agreement between the number of observed species and the total number of predicted species, confirming that the sampling method was appropriate and sampling complete. Correlations between the copepod community structure and measured parameters, obtained with Pearson correlation coefficient and principal component analysis (PCA) show different patterns. Presence of copepods was positively correlated with conductivity and dissolved organic carbon in Grotta di Roverè Mille, and with carbonate and nitrate in Pološka jama. In other caves not so high correlations were found between copepod species and measured parameters and between pairs of species. Investigations of the Alpine and Pre-alpine epikarst fauna can help to understand better the ecology of the epikarst fauna and its roles within the large range of different shallow subterranean habitats.
Ključne besede: Epikarst, Alpine and Pre-Alpine caves, percolating water, speleobiology, fauna.
Objavljeno: 31.08.2016; Ogledov: 1504; Prenosov: 99
.pdf Polno besedilo (2,00 MB)

29.
SINGLE NUCLEOTIDE POLYMORPHISMS OF THE CHROMOSOME SEGREGATION GENES INVOLVED IN THE DEVELOPMENT OF GASTRIC CANCER
Marija Rogar, 2016, doktorska disertacija

Opis: INTRODUCTION. Gastric cancer represents the fifth most frequently diagnosed cancer in the world. Despite numerous research studies, mechanisms leading to disease are poorly known and unclear. At the molecular level, many changes are involved in the development of gastric cancer, including malfunction of chromosome segregation genes. These abnormalities can lead to chromosomal instability (CIN). Segregation gene function can be affected by the low penetrance errors which include polymorphisms. AIM. The aim of this study is to examine the effect of selected polymorphisms in specific segregation genes on gastric cancer development. HYPOTHESIS. The study focused on exploring genotypes of selected polymorphisms in specific mitotic segregation genes. Those that differ significantly between the subjects and the healthy control population, may be associated with higher risk for developing gastric cancer or with certain clinical and histopathological characteristics, and may have effect on the survival of gastric cancer patients. METHODS. 30 polymorphisms in genes BUB1B, CASC5, ESPL1, PTTG1, SMC1A, TPX2, TTK and ZWINT were included in the study. Subjects were compared with the control group. Polymorphisms were determined using quantitative real-time PCR (qPCR), restriction fragment length polymorphism (RFLP) and DNA sequencing. RESULTS. The association between polymorphisms rs2277559 (BUB1B), rs2241666 (ZWINT), rs11858113 (CASC5) and rs11855334 (CASC5) and increased risk of developing gastric cancer in male population was determined. As concerning rs11855334, statistically significant difference was also observed in the genotype distribution between the whole population of subjects and controls. The association between the genotypes of polymorphisms (in gene BUB1B) rs2277559, rs2290551, rs1801376, rs1047130, rs1565866, rs2277560 and Lauren classification was recognized. Genotypes of polymorphisms rs1801376 (BUB1B), rs11855334 (CASC5), rs2241666 (ZWINT), rs2910101 (PTTG1) and rs1047130 (BUB1B) are linked to different tumour differentiation grades. Survival analysis revealed association between the lymph node involvement and perineural invasion. Statistically higher frequencies of haplotypes G-A-G-T-G-G-A, G-G-A-G-A-A-G and A-G-G-T-A-G-A in gene BUB1B and of haplotypes A-A-A-C and C-C-G-T in gene ESPL1 were observed in gastric cancer patients, whereas haplotypes A-C-A-T and C-A-G-T in gene ESPL1 were significantly more frequent in the control group. Association with gastric cancer was not noted with other polymorphisms. CONCLUSIONS. The association between specific polymorphisms of selected chromosome segregation genes and gastric cancer was recognized. Findings could provide guidelines for further research and polymorphisms linked to gastric cancer could serve as potential diagnostic and prognostic biomarkers.
Ključne besede: gastric cancer, chromosomal instability, polymorphisms, segregation genes
Objavljeno: 27.07.2016; Ogledov: 1766; Prenosov: 122
.pdf Polno besedilo (2,87 MB)

30.
A REVERSE GENETIC SYSTEM FOR ROTAVIRUS
Giuditta De Lorenzo, 2016, doktorska disertacija

Opis: Studies on rotavirus biology have always suffered for the lack of a reverse genetics system applicable to all genome segments and independent from the use of helper virus. One of the main reasons proposed to explain the difficulties encountered is the poor expression of the transfected cDNAs. In this work we investigated the role of 5’UTRs in rotavirus cDNAs expression in un-infected cells. We showed that in the 5’UTR of several (but not all) genome segments of rotavirus is present a 5’-terminal inhibitory motif (IM = 5’-GGY(U/A)UY-3’) that, independently from the ORF or the 3’UTR, downregulates both transcription and translation of rotavirus cDNAs when expressed exploiting T7 polymerase-encoding recombinant vaccinia virus. We mapped two mutations (insertion of a G upstream 5’UTR and the U to A mutation of the fifth nucleotide of IM) that are capable of making the inhibitory motif non-functional restoring a satisfying yield of protein synthesis. After the analysis of all genome segment 5’UTR of SA11 strain, we created two distinct sets of mutated rotavirus genome segments containing non-functional IM. We planned to employ these sets in the development of a plasmid-based reverse genetics system that exploit vaccinia virus-encoded T7 polymerase. In an attempt to produce in vivo biotinylated Tripled-Layered Particles (TLPs), we created a recombinant VP4 fused to the Biotin Acceptor Peptide (BAP) that, when co-expressed with the biotin ligase BirA, is efficiently in vivo biotinylated. We exploited recombinant vaccinia virus system to achieve high levels of VP4-BAP and the enzyme BirA in cells. Upon infection with rotavirus, VP4-BAP would be incorporated in the newly forming viral particles. In addition, we constructed a full-length genome segment 4 cDNA encoding the recombinant VP4-BAP to be used with a VP4 temperature sensitive rotavirus in order to generate a recombinant rotavirus encoding VP4-BAP. Thus, the results obtained allowed us to design two distinct possible ways of generating recombinant rotavirus. The first was a genetic strategy for the incorporation of an exogenous genomic segment, with the characteristic of being helper-virus free and applicable to all genomic segments. This was the consequence of the description of an inhibitory motif present in many viral mRNAs and the identification of two mutations that abrogate its inhibitory activity allowing expression of viral proteins. The second focused on the expression of a recombinant viral structural protein expressed in virus-infected cells for the consequent incorporation into newly made virions.
Ključne besede: rotavirus, UTR, reverse genetics, T7 polymerase, in vivo biotinylation
Objavljeno: 08.06.2016; Ogledov: 1058; Prenosov: 96
.pdf Polno besedilo (2,87 MB)

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