Dissertations / Theses on the topic 'Conserved region'
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Severi, Emmanuele. "The role of the conserved carboxy-terminal region of the Escherichia coli ammonium channel AmtB." Thesis, University of East Anglia, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.426575.
Full textZhang, Xiaoyan. "Phosphorothioate hammerhead ribozymes targeting a conserved sequence in V3 loop region inhibit HIV-1 infection." Kyoto University, 1999. http://hdl.handle.net/2433/181687.
Full textEno-Ibanga, Cheryl K. "The analysis of a conserved RNA structure in the 3D polymerase encoding region of human parechovirus 1." Thesis, University of Essex, 2016. http://repository.essex.ac.uk/19097/.
Full textAhmed, Tina May. "Characterisation of T cells induced by candidate conserved region HIV-1 vaccines in healthy HIV-1/2 negative volunteers." Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:8d06f1f6-dd2a-4be1-b66c-936c5d006f38.
Full textWang, Yibing. "Study of cysteines in the stalk region of CD3 proteins : evolutionarily conserved residues critical for T cell development and function /." Connect to full text via ProQuest. Limited to UCD Anschutz Medical Campus, 2008.
Find full textTypescript. Includes bibliographical references (leaves 138-153). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;
Van, Vleet Eric. "From Passive to Active Community Conservation: A Study of Forest Governance in a Region of the Sierra Norte of Oaxaca, Mexico." FIU Digital Commons, 2013. http://digitalcommons.fiu.edu/etd/823.
Full textOlinski, Robert Piotr. "Evolutionary Analysis of the Insulin-Relaxin Gene Family from the Perspective of Gene and Genome Duplication Events." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7892.
Full textXu, Lin. "HIV-1 mucosal immunity : from infection to prevention : HIV-1 envelope gp41 conserved region P1 modulates the mucosal innate immune response and acts as a potential mucosal adjuvant The HIV-1 viral synapse signals human foreskin keratinocytes to secrete thymic stromal lymphopoietin facilitating HIV-1 foreskin entry By shaping the antigen binding site in IgA, the CH1α domain is crucial for HIV-1 protection in highly exposed sero-negative individuals The antigen HIV-1 envelope gp41 conserved region P1 can act as mucosal adjuvant by modulating the innate immune response." Thesis, Sorbonne Paris Cité, 2018. http://www.theses.fr/2018USPCB071.
Full textMucosal vaccination, especially intranasal administrated ones, has been considered to be ideal for protection from pathogens invading through mucosal sites. However, the lack of specific adjuvant and insufficient acknowledgement of nasal immune system limits the development of vaccine. P1, a conserved region of gp41 envelope glycoprotein, was recently developed into a prophylactic HIV-1 vaccine immunized via both the intramuscular and intranasal routes. It showed high efficiency in pre-clinical and phase I clinical trial due to induction of P1 specific mucosal IgA with transcytosis blocking activity and IgG inducing antibody dependent cell cytotoxicity. In this study, we characterized the immunological mechanism underneath P1-vaccine in nasal mucosa. Firstly, we demonstrated that P1 initiate immune responses by inducing nasal epithelial cells to secret the Th2 cytokine Thymic Stromal LymphoPoietin (TSLP). TSLP has been reported to be a strong mucosal adjuvant, and its receptor TSLP-R plays a critical role in IgA response. We showed that P1 induce TSLP expression via the interaction with galactosyl ceramide, the receptor of HIV-1 mucosal entry. Furthermore, we identified Calcineurin/NFAT signaling pathway and microRNA-4485 as important players in the regulation of TSLP production induced by P1. Secondly, we showed that P1 modulates innate immune responses by activate dendritic cells (DCs). P1 stimulation results in enhanced expression of costimulatory molecules on DCs. Furthermore, the secretion of IL-6, IL-10 were increased, while IFN-γ was reduced, indicating that P1 activated DCs polarize into a Th2 and IgA prone phenotype. In addition, IL-8, CCL20, CCL22 were produced indicating a capacity at recruiting immune cells to mucosal surface for initiation of an adaptive immune response. MMP-9 was also produced allowing degradation of the extracellular matrix and facilitating the migration of immune cells out of the mucosa. Stricingly, a TSLP autocrine loop was observed as P1 induced DCs to secret TSLP and meanwhile, enhanced DC expression of TSLP-R. Finally, P1 activated DCs enhanced the proliferation of CD4+ T cells. In conclusion, we demonstrated that P1 is a multi-functional protein with a great interest for vaccine development, not only as an antigen for vaccine candidate, but also as a potential adjuvant that can be combined to other mucosal vaccines
Dantas, M?rcia Danielle de Ara?jo. "Estudo do genoma do v?rus causador da mionecrose infecciosa em camar?es e desenvolvimento de m?todos para detec??o de polimorfismos." Universidade Federal do Rio Grande do Norte, 2014. http://repositorio.ufrn.br:8080/jspui/handle/123456789/12630.
Full textCoordena??o de Aperfei?oamento de Pessoal de N?vel Superior
Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites
A carcinicultura ? uma das atividades que mais contribui para o crescimento da aquicultura mundial. Entretanto, esta atividade vem sofrendo perdas econ?micas significativas devido ao surgimento de doen?as virais como a Mionecrose Infecciosa (IMN). A IMN j? est? disseminada em toda regi?o Nordeste do Brasil e atingiu outros pa?ses como Indon?sia, Tail?ndia e China. O principal sintoma da doen?a ? a mionecrose, que consiste na necrose dos m?sculos estriados do abd?men e do cefalot?rax do camar?o. A IMN ? causada pelo v?rus da mionecrose infecciosa (IMNV), um v?rus n?o envelopado que apresenta protrus?es ao longo de seu caps?deo. O genoma viral ? formado por uma ?nica mol?cula de RNA dupla fita e possui duas Open Reading Frames (ORFs). A ORF1 codifica a prote?na principal do caps?deo (MCP) e uma poss?vel prote?na de liga??o a RNA (RBP). A ORF2 codifica uma prov?vel RNA polimerase dependente de RNA (RdRp) e classifica o IMNV dentro da fam?lia Totiviridae. Assim, o objetivo desse estudo foi estudar o genoma completo do IMNV e as prote?nas codificadas no intuito de desenvolver um sistema que identificasse diferentes isolados do v?rus com base na presen?a de polimorfismos. A rela??o filogen?tica entre alguns totiv?rus foi investigada e mostrou um novo grupo para o IMNV dentro da fam?lia Totiviridae. Dois novos genomas foram sequenciados, analisados e comparados a outros dois genomas j? depositados no GenBank. Os novos genomas foram mais semelhantes entre si do que com aqueles j? descritos. Regi?es vari?veis e conservadas do genoma foram identificadas atrav?s de gr?ficos de similaridade e alinhamentos utilizando as quatro sequ?ncias do IMNV. Esta an?lise possibilitou o mapeamento de s?tios polim?rficos e revelou que a regi?o mais vari?vel do genoma se encontra na primeira metade da ORF1 e coincide com as regi?es que possivelmente codificam a protrus?o viral, enquanto que as regi?es mais est?veis se encontraram em dom?nios conservados de prote?nas que interagem com o RNA. Al?m disso, estruturas secund?rias foram preditas para todas as prote?nas empregando diversos softwares e modelos estruturais proteicos foram calculados usando modelagens por threading e simula??es ab initio. A partir dessas an?lises foi poss?vel observar que as prote?nas do IMNV possuem motivos e formas similares ?s prote?nas de outros totiv?rus, e novas poss?veis fun??es proteicas foram propostas. O estudo do genoma e das prote?nas foi essencial para o desenvolvimento de um sistema de detec??o baseado em PCR capaz de discriminar os quatro isolados do IMNV com base na presen?a de s?tios polim?rficos
Ramirez, Christina J. "BRCA genes : conserved regions and the potential effect of missense changes /." Thesis, Connect to this title online; UW restricted, 2005. http://hdl.handle.net/1773/5052.
Full textBaek, Daehyun. "Characterization of evolutionarily conserved mammalian alternative splicing and alternative promoters /." Thesis, Connect to this title online; UW restricted, 2007. http://hdl.handle.net/1773/8036.
Full textChoudhuri, Jomuna Veronica. "Bioinformatics approaches to large scale genome comparison, including the identification of conserved noncoding regions." [S.l. : s.n.], 2003. http://deposit.ddb.de/cgi-bin/dokserv?idn=968573630.
Full textMoffet, Matthew Durwin. "Identifying regions of conserved synteny between pea (Pisum spp.), lentil (Lens spp.), and bean (Phaseolus spp.)." Thesis, Montana State University, 2006. http://etd.lib.montana.edu/etd/2006/moffet/MoffetM1206.pdf.
Full textCampanera, Reig Mireia. "¿Para quién se conserva la laguna Jacinto? Conflictividad socioambiental en el Bajo Marañón, Amazonía peruana." Doctoral thesis, Universitat de Barcelona, 2016. http://hdl.handle.net/10803/398394.
Full textA few days of fishing in 2009 reveal a latent conflict in the Jacinto lagoon (Pacaya Samiria National Reserve, Peru), raising different ways of conceiving, being, and intervening in the protected area and the lagoon. This situation is the consequence of the contradictions and dilemmas of two socio-environmental models: the sustainable development model and the local model of 'cuidado'. This research identifies and analyzes every agent involved in the fishing activity. First of all, the state institutions are the Direction of the Pacaya Samiria National Reserve and the Regional Department of Production. Second, the Spanish International Agency for Cooperation and Development (AECID). The first and the last one are a good example of the sustainable development agents. At the local level, there are three agents: the citizens of the San Jacinto community, the fishers of the organization called ‘Tigres Negros', and the Snake of the lake (a non-human being with recognized intention, from the local perspective). The conclusion shows that each agent has its relations, representations, and agenda around the lagoon. Thus, every agent takes its particular and complex position in this heterogeneous aquatic space. With a historical, political and cultural perspective, this doctoral dissertation analyzes the aquatic and fishery culture of San Jacinto and the low Marañón area. This case study contributes to understanding the complexity of the latent social and environmental conflict within the Pacaya Samiria National Reserve.
GUYOT, JEAN-BAPTISTE. "Mutagenese dirigee des regions conservees de l'adenine adn methylase d'e. Coli (dam)." Paris 6, 1994. http://www.theses.fr/1994PA066141.
Full textFERNANDES, MARIE. "Etude comparative de deux regions chromosomiques conservees au cours de l'evolution : la bande q13 du chromosome 11 humain et la region pericentromerique du chromosome 19 de la souris." Aix-Marseille 2, 1997. http://www.theses.fr/1997AIX22059.
Full textPereira, Ryan Apolinario. "FUNCTIONAL ANALYSIS OF TWO CONSERVED REGIONS OF ESCHERICHIA COLI ELONGATION FACTOR G AS STUDIED BY SITE-DIRECTED MUTAGENESIS." The Ohio State University, 2002. http://rave.ohiolink.edu/etdc/view?acc_num=osu1039030876.
Full textPereira, Ryan A. "Functional analysis of two conserved regions of Escherichia coli elongation factor G as studied by site-directed mutagenesis /." The Ohio State University, 2002. http://rave.ohiolink.edu/etdc/view?acc_num=osu1486549482669521.
Full textJACQUET, PARNAUDEAU MARIE-ANGE. "Etude comparative de promoteurs d'e. Coli naturels et mutes dans les regions non conservees." Paris 7, 1987. http://www.theses.fr/1987PA077120.
Full textHing, Benjamin. "Investigating differential regulation of BDNF promoter IV activity by upstream polymorphic evolutionary conserved regions : implications for mood disorders and cognitive disfunction." Thesis, University of Aberdeen, 2011. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=185597.
Full textDanchin, Etienne Gaëtan Jacques. "Reconstruction of ancestral genomic regions by comparative analysis of evolutionary conserved syntenies : Towards reconstructing the genome of the ancestor of all Bilaterian species (Urbilateria)." Aix-Marseille 2, 2004. http://www.theses.fr/2004AIX22070.
Full textBarrantes, Dávila Samuel Alexander, and Samillán José Alexander Vidaurre. "Proyecto de preinversión para la instalación de una planta de procesamiento y comercialización de conservas de pescado en la región Lambayeque." Bachelor's thesis, Universidad Católica Santo Toribio de Mogrovejo, 2018. http://tesis.usat.edu.pe/handle/usat/1042.
Full textTesis
Salazar, Rondon Maria Claudia [Verfasser], Jane E. [Gutachter] Parker, and Alga [Gutachter] Zuccaro. "Role of evolutionary conserved MAP kinase C-terminal regions in transcriptional activation in Arabidopsis thaliana / Maria Claudia Salazar Rondon ; Gutachter: Jane E. Parker, Alga Zuccaro." Köln : Universitäts- und Stadtbibliothek Köln, 2019. http://d-nb.info/1213023629/34.
Full textGolin, Raíssa Ochôa. "Estudo da Evolução da Protease 3c de Picornaviridae e Vírus Picorna-like Através da sua Sequência Proteica e Domínios Conservados." Universidade Federal do Pampa, 2014. http://dspace.unipampa.edu.br:8080/xmlui/handle/riu/213.
Full textMade available in DSpace on 2015-05-07T22:43:52Z (GMT). No. of bitstreams: 1 126110038.pdf: 2182930 bytes, checksum: 1d0e56324e775d985abbaf5cea8f4038 (MD5) Previous issue date: 2014-03-21
As abelhas apresentam uma combinação de características individuais e ainda de cooperação animal não encontrada no restante do reino animal. São insetos sociais e participam da polinização de diversas plantas que fornecem alimento para o homem. No Brasil com a africanização das abelhas, essas tornaram-se altamente produtoras e enxameadoras, o que vem tornando o país uma potência na produção de mel e outros produtos originados da atividade apícola. Muitas doenças podem afetar as abelhas, dentre elas muitas causadas por vírus. Controlar as infecções virais é essencial para a manutenção ecológica das abelhas e da produção apícola. Ao mesmo tempo, existe a possibilidade de relacionar vírus que infectam humanos com os vírus que infectam as abelhas, visto que os tratamentos utilizados para os seres humanos poderiam ser utilizados em colméias e, ao mesmo tempo, utilizar as abelhas como modelo de estudo para o desenvolvimento de novos antivirais. Na busca por um ponto em comum analisamos filogeneticamente a protease 3C, que ocorre nos vírus da super-família Picorna-like, onde encontram-se os vírus que parasitam as abelhas. Essa protease tem a capacidade de clivar a poliproteína viral nas proteínas maduras do vírus e ainda causar a degradação proteolítica das proteínas do hospedeiro. Até hoje não foi encontrada uma forma de utilizar a protease 3C em estudos filogenéticos pois existe muita divergência das suas sequências entre os vírus. O objetivo dessa pesquisa foi identificar uma forma de analisar filogeneticamente a protease 3C. As sequências da protease 3C e da RdRp de 55 vírus foram coletadas do NCBI ( National Center of Biotechnology Information) e submetidas ao MEME ( Multiple Em for Motif Elicitation), onde foram obtidos quatro sítios conservados. Após foi realizada a análise filogenética dos sítios conservados por Máxima Verossimilhança e análise da distância entre os sítios por parcimônia e ainda foi construída uma árvore com base na RdRp. As árvores dos sítios 1 e 2 apresentaram uma melhor robustez estatística e agrupamento dos vírus. Essas regiões conservadas da protease 3C-Pro podem ser o início para estabelecermos uma relação entre as proteases dos picornavírus e vírus picorna-like na busca da compreensão do seu mecanismo de infecção viral e também uma alternativa de estudo para outras sequências com alta variabilidade. O uso dos domínios 1 e 2 proporcionou a árvore com maior robustez apresentada até o dia de hoje para esta proteína viral.
The bees have a combination of individual features and animal cooperation is not yet found in the rest of the animal kingdom . They are social insects and participate in the pollination of many plants that provide food for man . In Brazil with the africanization of bees , these have become highly producing and swarm , which is making the country a power in the production of honey and other products derived from beekeeping . Many diseases can affect bees , among them many caused by viruses . Control viral infections is essential for ecological maintenance of bees and beekeeping . At the same time , it is possible to relate viruses that infect humans and viruses that infect the bees , whereas the treatments for humans could be used in beehives and at the same time using the bees as a model for development of new antiviral agents. In the search for a common point analyzed phylogenetically 3C protease , which occurs in the superfamily Picorna -like, which are viruses that parasitize bees virus. This protease is capable of cleaving the polyprotein, the mature viral proteins and viruses also cause the proteolytic degradation of host proteins . Until today there a way to use the 3C protease was found in phylogenetic studies because there is much divergence of their sequences between virus.The objective of this research was to identify a way to analyze phylogenetically 3C protease . The 3C protease and RdRp sequences of 55 viruses were collected from the National Center for Biotechnology Information , submitted to MEME , where four conserved sites were obtained . Upon phylogenetic analysis of the conserved sites and by maximum likelihood analysis of the distance between sites by parsimony was performed and was still a tree constructed based on the RdRp . Trees of sites 1 and 2 had a better statistical robustness and clustering of virus. These conserved regions of the 3C protease - Pro may be the start to establish a relationship between the proteases of the picornavirus and Picorna-like viruses in the quest to understand the mechanism of viral infection and also an alternative study for other sequences with high variability . The use of domains 1 and 2 provided the tree with greater robustness displayed until today for this viral protein.
Cabral, Byrne Pablo Enrique, and Pavlich Pablo Antonio José Arnillas. "Analizar la viabilidad de exportar alcachofa fresca congelada, corazones y fondos en conserva, desde la Región Junín, al mercado norteamericano." Master's thesis, Universidad Peruana de Ciencias Aplicadas (UPC), 2013. http://hdl.handle.net/10757/273834.
Full textTesis
MARIE-JEANNE, TORDO VERONIQUE. "Region 5 du genome du virus y de la pomme de terre : etude par sequencage de son polymorphisme et des motifs conserves. contribution a l'etude de la proteine p1." Paris 11, 1993. http://www.theses.fr/1993PA112434.
Full textGalliot, Sonia. "A la recherche de nouvelles AgNORs: une famille de protéines nucléolaires conservées et marqueurs potentiels du cancers." Doctoral thesis, Universite Libre de Bruxelles, 2010. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/210190.
Full text1-identifier des protéines AgNORs chez la levure
2-caractériser les propriétés physico-fonctionnelles et physico-chimiques de ces protéines AgNORs.
3-utiliser ces caractéristiques physico-chimiques pour rechercher de nouvelles AgNORs humaines, spécifiques de processus de cancérisation et potentiellement utilisables comme marqueurs tumoraux./The nucleolus is a subnuclear compartment that organized around ribosomal gene (rDNA) repeats NORs, which encode for ribosomal RNA. A peculiar group of acidic proteins which are highly argyrophilic are also localized at the same sites as NORs, thus allowing NORs to be very clearly and rapidly visualized by silver nitrate staining procedures. However, if three human argyrophilic proteins, UBF, C23 (nucleolin) and B23 (nucleophosmin), have been associated for staining of NOR, the exact number of AgNOR proteins and their intrinsic biochemical feature are unclear. Here, we have performed an heterologous screen in a genetically tractable eukaryotic organism (budding yeast) for the identification of novel AgNOR proteins and in vitro characterized an intrinsic feature that underlies silver binding and offers a strong predictive value for the identification of novel human AgNOR proteins.
Doctorat en Sciences
info:eu-repo/semantics/nonPublished
Pican?o, Jos? Reinaldo Alves. "Desenvolvimento, sustentabilidade e conserva??o da biodiversidade na amaz?nia: a produ??o familiar agroextrativista em ?reas protegidas no sul do amap?" Universidade Federal do Rio Grande do Norte, 2010. http://repositorio.ufrn.br:8080/jspui/handle/123456789/13731.
Full textThe establishment of Extractivism and Sustainable Development Reserves comes from an amazon forestry people resistance initiative. It means an option of natural resources management as protected areas for agroextractivism purposes. According to the institutional point of view, these lands, called Conservation Unity for Sustainable Exploration, belong to the government which grants the usufruct rights to the agroextractivist families under a sharing territory administration agreement among government and rural communities. The main roles of these lands are both: to improve the dwellers wellbeing, and protecting the local biodiversity. Additionally, they also represent the start of this thesis theme entitled Development, sustainability, and biodiversity conservation in the Amazon region: the use of protected areas for agroextractivism domestic yield in south of Amap? state with the objective of analyzing the performance that each territory has been reaching in terms of the attributions proposed at the beginning, when they were created. Social, economics, and environment changes that occurred in the agroextractivist areas have been evaluated from two selected test sites, named Rio Cajari Extractivist Reserve and Rio Iratapuru Sustainable Reserve, both, localized in the south of Amap? state
A cria??o de Reservas Extrativistas e Reservas de Desenvolvimento Sustent?vel, surgem a partir do movimento de resist?ncia dos povos da floresta amaz?nica, e representa uma alternativa de gest?o dos recursos naturais sob a forma de ?reas protegidas destinadas ao agroextrativismo. Do ponto de vista institucional, esses espa?os territoriais s?o Unidades de Conserva??o de Uso Sustent?vel, pertencentes ao poder p?blico, que concede o direito de usufruto ?s fam?lias agroextrativistas, num processo gest?o compartilhada desses territ?rios entre o poder p?blico e as representa??es comunit?rias. Essas ?reas t?m duplo objetivo, promover a melhoria das condi??es de vida dos moradores e garantir a prote??o da biodiversidade local. Esses objetivos constituem o ponto de partida desta tese Desenvolvimento, sustentabilidade e conserva??o da biodiversidade na Amaz?nia: a produ??o familiar agroextrativista em ?reas protegidas no sul do Amap?, que busca analisar em que medida esses territ?rios est?o cumprindo com as finalidades para as quais foram criados. A pesquisa foi realizada na Reserva Extrativista do Rio Cajari e na Reserva de Desenvolvimento Sustent?vel do Rio Iratapuru, localizadas no sul do estado do Amap?, analisando as mudan?as sociais, econ?micas e ambientais, ocorridas nessas ?reas agroextrativistas
Silva, Sandra Cristina Patrício da. "O que o estado português quis conservar: a avaliação e aquisição de documentos de arquivo em Portugal nos séculos XIX e XX." Master's thesis, Universidade de Évora, 2011. http://hdl.handle.net/10174/14829.
Full textPark, Jung Hee. "Crystal structure of ligand-free G-protein-coupled receptor opsin." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2010. http://dx.doi.org/10.18452/16049.
Full textRhodopsin as the visual pigment in photoreceptor cells is one of the most actively studied GPCRs. It consists of the apoprotein opsin and the inverse agonist, 11-cis-retinal. The inactivating ligand is bound in the seven-transmembrane helix (TM) bundle and cis/trans-isomerized by light to activate the receptor. The active receptor state is capable of catalyzing nucleotide exchange in the G protein and decays within minutes into opsin and all-trans-retinal. The visual pigment is then restored by reloading opsin with new 11-cis-retinal. In the present work, the successful crystallization of native opsin from bovine retinal rod cells and determination of the protein structure to 2.9 Å resolution is presented. Compared with the known structure of inactive rhodopsin, opsin displays prominent structural changes in the conserved E(D)RY and NPxxY(x)5,6F regions and TM5-TM7. At the cytoplasmic side, TM6 is tilted outwards by 6-7 Å, whereas the helix structure of TM5 is more elongated and close to TM6. These structural changes, of which some are attributed to an active GPCR state, reorganize the empty retinal binding pocket to disclose two openings for exit and entry of retinal. The opsin structure thus sheds new light on binding of hydrophobic ligands to GPCRs, GPCR activation and signal transfer to the G protein.
Gerhardt, Cleyton Henrique. "Pesquisadores, popula??es locais e ?reas protegidas: entre a instabilidade dos lados e a multiplicidade estrutural das posi??es." Universidade Federal Rural do Rio de Janeiro, 2008. https://tede.ufrrj.br/jspui/handle/tede/731.
Full textFunda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro
The relationship between local populations and protected areas has been an extremely controversial issue. However, this divergences also extend to the scientific domain, mobilizing the attention of scientists, who are interested both in researching specific aspects and in interfering in public policies related to this issue. If there is indeed a general agreement among specialists, then it is about the fact that the discursive context of this issue is marked by dissension, polyphony and by fierce academic dialogues. Bearing this in mind, I observed this controversial world reflected in the relationships among the scientists who, by operating in the frontiers of political action and scientific research, got involved in this debate. The thesis is divided into two parts. In the first part, some interpretative similarities and dissimilarities between authors are analysed and described based on their publications. As I intend to show, antagonisms, oppositions, divergences, but also alliances, agreements and convergences generate, within a wider strength balance marked by different identity/alterity levels, a permanently unstable and inconsistent structural environment. The second part is also divided into two distinct chapters. In the first chapter, I worked on fragments of different social trajectories and life experiences reported by 33 researchers I had the opportunity to interview, which allowed an insight into an extremely heterogeneous set regarding the paths they followed. In the last chapter, I present their observations, reflections, assessments and criticism on some aspects related to policies that target local populations and protected areas.
A rela??o entre popula??es locais e ?reas protegidas ? tema hoje extremamente controvertido no ?mbito cient?fico, mobilizando a aten??o de cientistas interessados tanto em pesquisar quest?es espec?ficas como em interferir em pol?ticas p?blicas a ele direcionadas. Se h? um consenso entre especialistas, ? que estamos diante de um contexto discursivo caracterizado pelo dissenso, pela polifonia e por ?cidos di?logos acad?micos. Diante disso, passei a observar esse universo controvertido que marca a rela??o entre cientistas que, atuando nas fronteiras da a??o pol?tica e da pesquisa cient?fica, se envolveram com este debate. Como tentei mostrar, oposi??es, diverg?ncias, mas, tamb?m, alian?as e converg?ncias geram, dentro de um equil?brio de for?as marcado por planos de identidade/alteridade distintos, um ambiente estrutural inst?vel. Dividi a tese em duas partes. Na primeira, problematizo e descrevo, a partir das suas respectivas publica??es cient?ficas, encontros e desencontros interpretativos protagonizados pelos autores. A segunda parte traz dois cap?tulos. No primeiro, trabalhei com fragmentos de diferentes trajet?rias sociais e experi?ncias de vida relatadas por 33 pesquisadores que tive a oportunidade de entrevistar, o que permitiu visualizar um quadro extremamente heterog?neo quanto ?s trilhas por eles percorridas. No ?ltimo cap?tulo apresento suas observa??es, reflex?es, avalia??es e cr?ticas sobre alguns aspectos relacionados ?s pol?ticas direcionadas ?s popula??es locais e ?reas protegidas. Ao final, al?m de apontar um inconveniente ?tico vinculado ? abordagem do estudo que realizei, reconecto alguns aspectos discutidos ao longo da tese com vistas a indicar o car?ter problem?tico que h? por tr?s da cristaliza??o de controv?rsias cient?ficas fortemente politizadas.
Lin, Yu-Shing, and 林育星. "Enzyme Class Prediction via Mining Conserved Region in Sequence and Structure." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/58424169935748829904.
Full text國立臺灣大學
資訊工程學研究所
94
The active sites and binding sites of the protein structure are the main working areas of the functional proteins. Based on this assessment, the qualitative and quantitative analyses of the variant characteristics of the active site and binding site are the first steps towards predicting protein functionalities. These characteristics include conformation, bio-chemical property etc. Present methods to predict protein function mostly exploit known protein-ligand complex to conduct factitious or automatic structure analysis to obtain the Motif of surrounded atoms of binding site. However, this thesis is providing a different point of view. Our assumption is though the protein function is decided by the location of active site or binding site, there should be some local conserved regions between the proteins that possess same function. In other words, these local conserved regions might not be the surrounded amino acids of binding site, but it indeed affects the structure and the way of working for active site or binding site. To describe the framework of these areas, it can’t be decided only by a single sequence or the structure analysis. What it needs is a further understanding of the relationship between sequence and structure, and takes it into consideration. To reach this goal, we developed the following steps. First, circle each residue around 10Å as a spheroid to represent a local region. Second, use each spheroid to conduct sequence and structure analysis. Third, with enzyme data bank, we can identify the belonging local conserved region of protein through same category of protein. In the future, further discussion about the relationships between theses local conserved region and the functions should be investigated.
Duque, Hernando. "Phenotypic characterization of three phylogenetically conserved stemloops in the Mengovirus 3 ́untranslated region." 1998. http://catalog.hathitrust.org/api/volumes/oclc/40944597.html.
Full textTypescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 146-156).
Huang, Liang-Yin, and 黃亮尹. "Identification and Characterization of the Conserved Region 1 of Bacillus subtilis sigmaA Factor." Thesis, 2001. http://ndltd.ncl.edu.tw/handle/39565274079948701067.
Full text國立中興大學
生物化學研究所
89
The sigma subunit of prokaryotic RNA polymerase is essential for promoter recognition and initiation of transcription. Our previous study has shown that the deletion mutant sigmaA factors, SND73- and SND94-sigmaA, are active in transcription after reconstitution with core RNA polymerase; however, SND129-sigmaA is inactive due to the loss of promoter-binding activity of the mutant sigmaA-RNA polymerase. The present study is aimed to explore how much amino acid residues in the N-terminal region of sigmaA are unnecessary for its full in vitro activity, and why further deletion from this amino acid position would cause the loss of transcription activity of the mutant sigmaA factor. Besides, we hope to obtain a series of functional sigmaA factors, which would enable us to study the crystal structure or the structural and functional relationship of sigmaA factor in the future. To fulfill this goal, 25 mutant sigmaA factors, with deletion at the N-terminus and spanning amino acid no. 74 to 127 of sigmaA, were constructed and purified. The functional properties of these mutant sigmaA factors were analyzed in vitro. Our data revealed that deletion of more than 103 amino acids at the N-terminus of sigmaA have resulted in a significant loss of the transcription activity of the reconstituted mutant sigmaA-RNA polymerases. Gel retardation assays indicated that the inactivation of transcription of these mutant sigmaA-RNA polymerases was due to the reduction of promoter-binding activity of the reconstituted holoenzyme. Since no drastic change in the protein conformation was observed among the free SND100-, SND103-, SND106- and SND109-sigmaA factors, and that a more extended deletion at the N-terminus of sigmaA (up to 129 amino acid deletion) would increase the core-binding activity of mutant sigmaA factors, I propose that sigmaA factors with more than 103 amino acids being deleted at the N-terminus do not possess functional conformation when they are associated with core RNA polymerase.
Lakowski, Jörn. "Analysis of an evolutionarily conserved distant regulatory region downstream of the Pax6 gene." 2003. http://purl.galileo.usg.edu/uga%5Fetd/lakowski%5Fjorn%5F200308%5Fms.
Full textMao, Wei-Chun, and 毛瑋俊. "Structural and functional distinctions of the non-major conserved region in Aeromonas caviae D1 chitinase." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/13556904675155049098.
Full text國立海洋大學
食品科學系
91
N-acetylchitooligosaccharides have already been used in many fields and it’s also valuable. Our laboratory had made efforts in this field for several years. We had cloned high chitinolytic activity chitinase gene from Aeromonas caviae D1, and we had established the optimum condition of induction-expression and the production of N-acetylchitooligosaccharides. When we were proceeding the alignment of the amino acid sequence of A. caviae D1 ChiA, we found that it’s highly conserved with the putative active site of Alteromonas sp. O-7 ChiA, Serratia marcescens ChiA, ChiB and Bacillus circulans WL-12 ChiA1, ChiD. After we had proceeded the point mutation in the chitinase conserved region of A. caviae D1 ChiA, we found the significant decrease of N-acetylchitobiase and N-acetylchitotriase activity, but it’s not totally destructed. Therefore we presumed other regions besides the conserved one must have its contribution. In the study, we cloned the non-major conserved region chi300 and the chitin binding domain chBD in the A. caviae D1 ChiA. They were expressed in the E. coli BL21 (DE3) pLysS using the pET-20b(+) expression system. The His-Tag-affinity-purified recombinant Chi300 and ChBD had molecular mass about 38.4 and 15.5 kDa. When we were proceeding the activity analysis of Chi300 and ChBD, we found that they don’t have activity toward lower oligomerized substrates. The N-acetylchitobiase and N-acetylchitotriase activity of G561 are about 41.79% and 44.65% of ChiA, respectively. It showed the non-major conserved region Chi300 could contribute to the whole enzyme hydrolysis toward lower oligomerized substrates. When we added Chi300 and ChBD to the ChiA or G561, we didn’t find the significant effect of hydrolysis in lower oligomerized substrates. In the test of substrate binding specificity, neither Chi300 nor ChBD could have satisfying binding ability. We presumed during the protein folding, Chi300 and ChBD had already lost the normal conformation in ChiA. It could be caused by the amino residues besides the ChBD blocked the angle between two planes during the protein folding. So, it destroyed the binding ability.
Shaffer, Robert. "Role of a highly conserved region of the NF-kappaB essential modulator in its scaffolding function." Thesis, 2018. https://hdl.handle.net/2144/34456.
Full text(9143657), Phillip S. Rushton. "Structure of the Plant-Conserved Region of Cellulose Synthase and Its Interactions with the Catalytic Core." Thesis, 2020.
Find full textThe processive plant cellulose synthase (CESA) synthesizes (1→4)-β-D-glucans. CESAs assemble into a six-fold symmetrical cellulose synthase complex (CSC), with an unknown symmetry and number of CESA isomers. The CSC synthesizes a cellulose microfibril as the fundamental scaffolding unit of the plant cell wall. CESAs are approximately 110 kDa glycosyltransferases with an N-terminal RING-type zinc finger domain (ZnF), seven transmembrane α-helices (TMHs) and a cytoplasmic catalytic domain (CatD). In the CatD, the uridine diphosphate glucose (UDP-Glc) substrate is synthesized into (1→4)-β-D-glucans. The ZnF is likely to facilitate dimers in the CSC. Recombinant class-specific region (CSR), a plant specific insertion to the C-terminal end of the CatD is also known to form dimers in vitro. The CSR sequence is the primary source of distinction between CESA isoforms and class structure. Also within the CESA CatD is a 125-amino acid insertion known as the plant-conserved region (P-CR), whose molecular structure was unknown. The function of the P-CR is still unclear, especially in the context of complete CESA and CSC structures. Thus, one major knowledge gap is understanding how multimeric CSCs synthesize multiple chains of (1→4)-β-D-glucans that coalesce to form microfibrils. The specific number of CESAs in a CSC and how interactions of individual CESA isoforms contribute to the CSC are not known. Elucidating the structure-function relationships of the P-CR domain, and with the consideration of the ability of CSR and ZnF domains to dimerize, it is possible to more completely model the structure of the CSC.
Recombinantly expressed rice (Oryza sativa) secondary cell wall OsCESA8 P-CR domain purifies as a monomer and shows distinct α-helical secondary structure by circular dichroism analysis. A molecular envelope of the P-CR was derived by small angle X-ray scattering (SAXS). The P-CR was crystallized and structure solved to 2.4 Å resolution revealing an anti-parallel coiled-coiled domain. Connecting the coiled-coil α-helices is an ordered loop that bends back towards the coiled-coils. The P-CR crystal structure fits the molecular envelope derived by SAXS, which in turn fits into the CatD molecular envelope. The best fit places the P-CR between the membrane and substrate entry portal. In depth analysis of structural similarity to other proteins, and 3D-surface structure of the P-CR, leads to hypotheses that it could function in protein-protein interactions as a dimer, trimer or tetramer in the CSC, that it could form protein-protein interactions with CESA-interacting proteins, and/or modulate substrate entry through its N- and/or C-terminus. From modeling, hypothetically important residues within the P-CR or related to the P-CR through potential protein contacts were mutated in Arabidopsis thaliana AtCESA1 constructs. These constructs were expressed in the temperature-sensitive radial swelling (rsw) rsw1-1 mutant of AtCESA1 to test for complementation of growth phenotypes at restrictive temperatures. Preliminary experiments indicate that some mutated CESA1 sequences fail to complement the rsw1-1 phenotype, suggesting that specific functions of individual amino can be tested using this system.
Jian, Ming Zhu, and 簡銘助. "Functional analysis of a small RNA generated from a highly conserved region of the Streptococcus parasanguinis FW213 chromosome." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/77ra2h.
Full textMannam, Praveen. "Immune response and protection against Streptococcus pyogenes after vaccination with Lactococcus lactis that expresses conserved region of M6 protein." Thesis, 2003. http://hdl.handle.net/1957/30816.
Full textGraduation date: 2004
Martínez-Lumbreras, S., E. M. Krysztofinska, A. Thapaliya, A. Spilotros, D. Matak-Vinkovic, E. Salvadori, P. Roboti, et al. "Structural complexity of the co-chaperone SGTA: a conserved C-terminal region is implicated in dimerization and substrate quality control." 2018. http://hdl.handle.net/10454/17888.
Full textProtein quality control mechanisms are essential for cell health and involve delivery of proteins to specific cellular compartments for recycling or degradation. In particular, stray hydrophobic proteins are captured in the aqueous cytosol by a co-chaperone, the small glutamine-rich, tetratricopeptide repeat-containing protein alpha (SGTA), which facilitates the correct targeting of tail-anchored membrane proteins, as well as the sorting of membrane and secretory proteins that mislocalize to the cytosol and endoplasmic reticulum-associated degradation. Full-length SGTA has an unusual elongated dimeric structure that has, until now, evaded detailed structural analysis. The Cterminal region of SGTA plays a key role in binding a broad range of hydrophobic substrates, yet in contrast to the well-characterized N-terminal and TPR domains, there is a lack of structural information on the C-terminal domain. In this study, we present new insights into the conformation and organization of distinct domains of SGTA and show that the C-terminal domain possesses a conserved region essential for substrate processing in vivo. We show that the C-terminal domain region is characterized by α-helical propensity and an intrinsic ability to dimerize independently of the N-terminal domain. Based on the properties of different regions of SGTA that are revealed using cell biology, NMR, SAXS, Native MS, and EPR, we observe that its C-terminal domain can dimerize in the full-length protein and propose that this reflects a closed conformation of the substrate-binding domain. Our results provide novel insights into the structural complexity of SGTA and provide a new basis for mechanistic studies of substrate binding and release at the C-terminal region.
MRC New Investigator Research Grant: G0900936; BBSRC grants: BB/L006952/1 and BB/L006510/1; BBSRC grant: BB/N006267/1; Wellcome Trust Investigator Award in Science: 204957/Z/16/Z; BBSRC grant: BB/J014567/1
Thompson, Sunnie R. "Characterization of conserved sequences within the 3' untranslated region of messenger RNA that regulate translation through changes in poly (A) tail length." 1998. http://catalog.hathitrust.org/api/volumes/oclc/40308502.html.
Full textchen, chun-chin, and 陳秋錦. "Functional analysis of the small RNA derived from the highly conserved 3’-untranslated region of Japanese encephalitis virus in infected mammalian cells." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/22361143479344785083.
Full text國立東華大學
生物技術研究所
93
英文摘要 Japanese encephalitis virus (JEV) contains a single positive-strand RNA genome of 10,976 nucleotides in length and is not formally thought to generate subgenomic RNA molecules during replication. Previous studies in our lab have reported the abundant accumulation of a 3’-terminal 521- to 523-nucleotide genome fragment, representing the highly conserved region of the 3’-untranslated region, in JEV-infected cells (Journal of Virology 78:5133, 2004). To address a possible function of the small RNA during viral replication, several approaches were carried out. Systematic quantification of plus- and minus-stand viral RNA synthesis using Northern hybridization, RNase protection, and RT-Real-time PCR assays suggested that the presence of the small RNA may play a role in the limitation of minus-stand RNA synthesis. Results from Northern analysis reveals that a minus-strand complement of the small RNA is not found, but rather only a minus-strand RNA that is 2X genome size is found. To elucidate a possible function of the small RNA and its complementary sequence during viral replication, unit-length (i.e., 523-nt) plus- and minus-strand forms of the small RNA were separately transfected in virus-infected cells and the effects on plusand minus-strand accumulation were measured. By strand-specific Northern hybridization and RT-real-time PCR assays. Transfection of the plus-sense small RNA appeared not to affect plus-strand viral RNA accumulation. However, transfection of the minus-sense small RNA caused a change in the migration pattern of the normally observed 2X minus-strand RNA in that nearly equal amounts of 1X-sized minus-strand 3 RNA are now found. The effect of transfection of the small plus-strand RNA on minus-strand accumulation remains to be determined. These results suggest that features of the minus-strand RNA may play a regulatory role during RNA synthesis in vivo.
Van, der Sluis Rencia. "Investigation and characterisation of the genetic variation in the coding region of the glycine N-acyltransferase gene / Rencia van der Sluis." Thesis, 2015. http://hdl.handle.net/10394/15639.
Full textPhD (Biochemistry), North-West University, Potchefstroom Campus, 2015
Khare, Tarang [Verfasser]. "Influence of assisted reproductive technologies on imprinting centers and functional characterization of conserved elements in Beckwith Wiedemann syndrome (BWS) region / by Tarang Khare." 2007. http://d-nb.info/982693001/34.
Full textChang, Yu-Chia, and 張毓嘉. "Study on the Importance of the Conserved Octamer 5’-GGAAGAGC-3’ Located in the Viral 3’ Untranslated Region for the Translation of Coronaviruses." Thesis, 2019. http://ndltd.ncl.edu.tw/cgi-bin/gs32/gsweb.cgi/login?o=dnclcdr&s=id=%22107NCHU5628005%22.&searchmode=basic.
Full text國立中興大學
獸醫病理生物學研究所
107
The 3’ untranslated region (UTR) of betacoronavirus consists of four functional RNA secondary structures: bulged stem-loop (BSL), pseudoknot (PK), hypervariable region (HVR), and minimal region at 3’ terminus, and these structures have been demonstrated to be able to regulate coronavirus gene expression in cis. Within the hypervariable region (HVR), the sequence motif 5’-GGAAGAGC-3’, which is defined as octamer (oct), is almost universally conserved among coronaviruses, suggesting its potentially critical role in coronavirus biology; however, its function remains unknown. Previous study has shown that the oct is not essential for viral replication of murine hepatitis virus (MHV) in vitro; however, the HVR (including oct) deletion mutants affects MHV pathogenesis in infected mice. Since viral replication and pathogenicity are associated with viral proteins, it is possible that the oct may affect MHV pathogenicity through modulating viral translation. Therefore, we hypothesized that the conserved oct within the HVR plays a crucial role in coronavirus translation. For this aim, we constructed various oct or HVR mutants in both bovine coronavirus (BCoV) defective interfering (DI) RNA and MHV full-length genome to determine the effects of the mutations on translation in cells by Western blot and thus the function of oct on coronavirus translation. The results suggested that the oct has positive regulatory function for translation of the BCoV DI RNA, subgenomic RNA and the MHV full-length genome. The sequence specificity of oct and the HVR structure where oct is located are associated with coronavirus translation. In addition, other RNA elements located in the 3’ UTR, such as BSL and PK, may also be involved in the coronavirus translation. In this study, we for the first time demonstrate the important role of the universal octamer 5’-GGAAGAGC-3’ for coronavirus translation. In addition, we also provide the evidence that the other RNA elements in the viral 3’ UTR may participate in the translation mechanism of coronaviruses.
Uthaman, Yazhisai, and 葉意香. "Untranslatable tospoviral NSs fragment enhances the broad resistance conferred by L gene conserved region in transgenic plants with or without a selection marker gene." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/41454221085296715568.
Full text國立中興大學
植物病理學系所
103
ABSTRACT Tospovirus, the only plant-infecting viruses in the family Bunyaviridae, cause serious damages on various economic crops all over the world. Watermelon silver mottle virus (WSMoV) and Melon yellow spot virus (MYSV), the two members of the genus Tospovirus, are the major limiting factors for cucurbits cultivation in Taiwan. In our earlier studies, transgenic tomato lines carrying the conserved region containing the RNA- dependent RNA polymerase (RdRp) motifs within the L gene of WSMoV confer broad-spectrum resistance against different tospoviruses mediated by post-transcriptioned gene silencing (PTGS). In this study, the enhanced broad resistance against distinct tospoviruses, in transgenic tobacco plants conferred by the conserved regions of L, N and NSs genes of tospoviruses were generated. Furthermore, marker-free transgenic tobacco plants carrying the L,N and NSs conserved regions of WSMoV and conferring broad resistance to distinct tospoviruses were produced . Our marker-free approach was further extended to produce marker-free economically important crops such as melon (Cucumis melo L.) carrying the conserved regions of L,N and NSs genes of WSMoV. Chapter 1, “Literature review and research objective” describes the background informations, references and research objectives of this this study. Chapter 2, “Untranslatable tospoviral NSs fragment coupled with L conserved region enhances transgenic resistance against the homologous virus and a serologically unrelated tospovirus”. In this study, Nicotiana benthamiana transgenic lines carrying individual untranslatable constructs comprised of the conserved region of the L gene (denoted as L), the 5′ half of NSs coding sequence (NSs) or the antisense fragment of whole N coding sequence (N) of Watermelon silver mottle virus (WSMoV), individually or in combination, were generated. A total of 15 -17 transgenic Nicotiana benthamiana lines carrying individual transgenes were evaluated against the WSMoV and the serologically unrelated Tomato spotted wilt virus (TSWV). Among lines carrying single or chimeric transgenes, the level of resistance ranged from susceptible to completely resistant against WSMoV. From the lines carrying individual transgenes and highly resistant to WSMoV (56-63% of lines assayed), 30% of the L lines (3/10 lines assayed) and 11% of NSs lines (1/9 lines assayed) were highly resistant against TSWV. The chimeric transgenes provided higher degrees of resistance against WSMoV (80-88%), and the NSs fragment showed an additive effect to enhance the resistance to TSWV. Particularly, the chimeric transgenes with the triple combination of fragments, namely L/NSs/N or HpL/NSs/N (a hairpin construct), provided a higher degree of resistance (both 50%, with 7/14 lines assayed ) against TSWV. Our results indicate that the untranslatable NSs fragment is able to enhance the transgenic resistance conferred by the L conserved region. The better performance of L/NSs/N and HpL/NSs/N in transgenic N. benthamiana lines suggests their potential usefulness in generating high levels of enhanced transgenic resistance against serologically unrelated tospoviruses in agronomic crops. Chapter 3, “Marker-free homozygous transgenic tobacco plants with resistance against tospoviruses of different serogroups and serotype”. In this study, to develop selectable marker-free transgenic plants resistant to different tospoviruses, we adopted a co-transformation system. Agrobacterium harboring pBI2T-based binary vectors, each with two T-DNAs separately carrying the selection marker gene (SMG) nptII and the construct with the aforementioned L fragment, alone or in combination with the NSs and N fragments , were used for transformation of Nicotiana benthamiana plants. Transgenic lines (12-18 for each construct) with high degrees of resistance against two serologically unrelated tospoviruses, Watermelon silver mottle virus (WSMoV) and Tomato spotted wilt virus (TSWV), were obtained. R1 progenies of 7 self-pollinated highly resistant transgenic R0 lines were further analyzed for the segregation of the SMG and the viral construct by PCR. When the individuals of the R3 progeny of the marker-free R2 homozygous transgenic plant were challenged with members of different tospoviruses of WSMoV, TSWV and Impatiens necrotic spot virus ( INSV ), resistance levels of 90-100% were noticed, indicating that the marker-free R2 and R3 homozygous transgenic N. benthamiana plants confer high degrees of broad resistance against tospoviruses of different serogroups and serotype. This approach is currently being extended to commercially important crops such as tomato, watermelon and melon. Supplementary information, “Generation of transgenic melon plants with marker-free hairpin construct” . In this study, using the co-transformation method, Agrobacterium harboring pBI2T 1.5 binary vector with two T-DNAs carrying the nptII selection marker gene and the gene of interest HpL/NSs/N (constructed using conserved regions of tospoviral L (replicase), N (nucleocapsid) and NSs (gene silencing suppressor) genes of tospoviruses) was used for transformation of melon (cv.Silver light) plant. The regenerated marker-free transgenic 12 melon plant lines showed the presence of transgene and marker gene when checked through PCR. The marker-free transgenic melon plant lines will be challenged with different tospoviruses such as WSMoV and MYSV.
Peng, Jui-Chu, and 彭瑞菊. "Occurrence of thrips-borne viruses infecting cucurbits in Taiwan and generation of broad-spectrum resistance in transgenic plants conferred by the conserved region of L genes of tospoviruses." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/59418743980813354305.
Full text國立中興大學
植物病理學系所
99
Thrips-borne tospoviruses, belonging to the only plant-infecting genus Tospovirus in the family Bunyaviridae that cause severe damages in economic crops worldwide, are globally important. Watermelon silver mottle virus (WSMoV) and Melon yellow spot virus (MYSV), the members of the genus Tospovirus, are of the major threats for the cultivation of cucurbits in Taiwan. Melon (Cucumis melo L.) and watermelon (Citrullus lanatus Thunb.) are economically important crops in Taiwan. In this study, the occurrence of thrips-borne viruses infecting melon in Taiwan was investigated, and the broad-spectrum resistance in transgenic plants conferred by the conserved region of L genes of tospoviruses was generated. This dissertation is divided into four chapters as described below. Chapter 1, “ Literature review and research objectives” describes references relevant to this study and the objectives for the investigations. Chapter 2, “Occurrence of thrips-borne viruses infecting melon and watermelon in Taiwan” describing the incidence of virus infection in melon fields, a field survey was conducted from July 2007 to December 2009. The symptomatic tissues were collected from the principal cultivated areas in central and southern Taiwan. The anti-N protein-monoclonal antibodies (MAbs) and the N gene-specific primer pairs can be used to identify WSMoV and MYSV. The uncertain ELISA results for tospoviruses were further verified by reverse transcription-polymerase chain reaction (RT-PCR) using N gene-specific primer pairs for WSMoV and MYSV. Among 10,480 melon samples tested, 631 (6%) and 1,906 (18.2%) were found infected with WSMoV and MYSV, respectively, and only 17 mixed infections by both WSMoV and MYSV. Our results indicated that MYSV is the major tospovirus to invade melon in Taiwan. On the other hand, among 1,811 watermelon samples assayed, 22.4% and 9.2% samples were singly infected with WSMoV and MYSV, respectively, and 4 samples were infected with both viruses. Our results indicated that mixed infection with the two thrips-borne viruses is rare. Moreover, host preference for both viruses is different; WSMoV prevails on watermelon whereas MYSV is more widespread on melon. We conclude that MYSV has become a serious threat for watermelon and melon production in Taiwan and the possible control measures are discussed. Chapter 3, “Generation of broad-spectrum resistance in transgenic plants conferred by the conserved region of L genes of tospoviruses ”. In this investigation, the conserved region containing the RNA-dependent RNA polymerase (RdRp) motifs within the L gene of WSMoV, an important cucurbit-infecting tospovirus in Taiwan, was used as a transgene for transformation of Nicotiana benthamiana and Solanum esculentum mediated by Agrobacterium tumefaciens to generate broad-spectrum resistance to tospoviruses. Five different modified transgene constructs of the L gene conserved region, including WLm in a sense translatable orientation, WLmt and WLmts in non-translatable, WLmAs in antisense and WLmds in double-stranded inverted repeat, were used to evaluate broad-spectrum resistance in transgenic plants. A total of 46.7-70.0% transgenic N. benthamiana lines derived from these five transgenes showed resistance to WSMoV, and 35.7-100% of the WSMoV-resistant lines exhibited broad-spectrum resistance against four other serologically unrelated tospoviruses, including Tomato spotted wilt virus (TSWV), Groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV) and Peanut chlorotic fan-spot virus (PCFV). Northern analyses indicated that the broad-spectrum resistance is mediated by post transcriptional gene silencing (PTGS). To validate the L conserved region resistance in vegetable crops, we have transferred all the transgenes constructs in transgenic tomato plants and the results indicated that this L conserved region generate high levels of resistance against WSMoV and other distinct tospoviruses. This is the first report that using a single nucleotide fragment corresponding to the L gene conserved region as a novel transgenic approach to trigger broad-spectrum resistance for controlling different tospoviruses at the genus level. Chapter 4, “Broad-spectrum resistance to tospoviruses conferred by the marker-free transgene constructs containing the L gene conserved region of Watermelon silver mottle virus in tomato and melon”. Using the co-transformation method, the conserved region containing the RdRp motifs within L gene of WSMoV was constructed in a two T-DNAs binary vector and used to generate marker-free transgenic plants. Two constructs- MF-WLmAs and MF-3WLmds were evaluated for the broad-spectrum resistance. Our results preliminary indicated that the L gene conserved region is successfully used to generate marker-free transgenic tomato and melon lines conferring broad-spectrum resistance against different tospoviruses. The segregation and elimination of the selection marker nptII will be selected from the progenies after selfing.
OLIVEIRA, NOGUEIRA MARCELA CRISTINA. "Characterization of heterogeneous viral proteins by NMR methods: Human Adenovirus E1A and Human papillomavirus E7 proteins." Doctoral thesis, 2016. http://hdl.handle.net/2158/1074811.
Full textCheng, Yu-Ting, and 鄭宇婷. "The effect of mutation of the non-conserved cysteine residue in the C-terminal region of the triple-gene-block protein 2 on the infectivity of Bamboo mosaic virus." Thesis, 2012. http://ndltd.ncl.edu.tw/handle/42283062301698130740.
Full text國立中興大學
生物化學研究所
100
The TGBp1, TGBp2 and TGBp3 encoded by the triple gene block (TGB) of Bamboo mosaic virus (BaMV) are required for virus movement across the cells.. The TGBp2 is transmembrane protein which has two conserved Cys residues (Cys-109 and Cys112) at their C-terminal tails. Previous study showed that the Cys-to-Ala substitutions of these two conserved residues in TGBp2 make the cell-to-cell movement of BaMV relatively inefficient and the systemic movement of BaMV severely inhibited. However, there is also a non-conserved Cys residue (Cys-119) at the C-terminal tail of TGBp2. To investigate whether Cys-119 of TGBp2 has similar importance to virus movement as the conserved ones, four mutant BaMVs which have Cys-to-Ala substitutions at position 119 of TGBp2 were constructed. and inoculated onto the leaves of Chenopodium quinoa and Nicotiana benthamiana. However, no disease symptom and GFP fluorescence were observed in the inoculated leave. To understand the reason causing the phenomenon, the amino sequence in the junction of TGBp2 and TGBp3 was analyzed. The result indicated that replacement of Cys-119 the final codon of TGBp2, with Ala will change the initiation codon, Met, of TGBp3 into Ser and thus affects the translation or the movement function of TGBp3. To overcome this problem, the Cys-119 of TGBp2 was replaced with Trp, which maintains the Met initiation codon of TGBp3. The infectivities of the four mutant BaMVs containing Cys-119-Trp were all lower than that of the wild-type. In addition, the TGBp2 in the membrane fraction of both the wild-type and mutant BaMV-infected N. benthamiana was analyzed by Western blot. Dimeric form of TGBp2, albeit with a much lower level, was detected in the tissues infected with the BaMV containing triple Cys substitutions in TGBp2. These results suggest that the conserved or non-conserved Cys residues are not essential for the formation of TGBp2 homo-multimer and it is the hydrophobic α-helical region which is responsible for the formation of TGBp2 multimer.