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A lentiviral gene transfer vector for the treatment of cystic fibrosis airway disease / Maria Limberis.Limberis, Maria January 2002 (has links)
"16th September 2002." / Accompanying CD contains 2 MPEG clips with accompanying text, and a copy in PDF format of: Recovery of airway cystic fibrosis transmembrane conductance regulator function in mice with cystic fibrosis after single-dose lentivirus-mediated gene transfer / M. Limberis ... [et al.], published in Human gene therapy vol. 13 (2002). / Bibliography: leaves xxix-li. / xxvii, 213, li leaves : ill., plates (some col.) ; 30 cm. + 1 CD-ROM (4 3/4 in.) / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This thesis focuses on modulating the physical barriers of the airway epithelium with mild detergents, so as to enhance gene transfer by a HIV-1 based lentivirus vector in vivo. The efficiency of the gene transfer was evaluated in the nasal airway of C57B1/6 mice using the Lac Z marker gene. This demonstration of lentivirus-mediated in vivo recovery of CFTR function in CF airway epithelium illustrated the potential of combining a pre-conditioning of the airway surface with a simple and brief HIV-1 based gene transfer vector exposure to produce therapeutic gene expression in the intact airway. / Thesis (Ph.D.)--University of Adelaide, Dept. of Paediatrics, 2003
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Construction and characterization of adenoviral vectors expressing cytokines for cancer immunotherapy /Addison, Christina Lynn January 1997 (has links)
Thesis (Ph.D) -- McMaster University, 1997 / Includes bibliographical references Also available via World Wide Web.
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Development of vaccines and experimental models for chronic infections caused by the hepatitis C virus /Frelin, Lars, January 2004 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.
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The development of synthetic gene delivery systems /Brandén, Lars J., January 1900 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2001. / Härtill 5 uppsatser.
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A gene transfer system derived from human immunodeficiency virus type 1 (HIV-1) /Fuller, Maria. January 2001 (has links) (PDF)
Thesis (Ph.D.)--University of Adelaide, Dept. of Public Health, 2002. / Bibliography: p. 189-229.
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Lentivirus-mediated gene expression in corneal endotheliumParker, Douglas George Anthony, January 2007 (has links)
Thesis (Ph.D.)--Flinders University, School of Medicine, Dept. of Ophthalmology. / Typescript bound. Includes bibliographical references: (leaves 258-281) Also available online.
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The construction of an expression vector for the transformation of the grape chloroplast genomeRobson, Julia 12 1900 (has links)
Thesis (MSc)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: The genetic information of plants is found in the nucleus, the mitochondria, and the plastids. The
DNA of plastids is comprised of multiple copies of a double-stranded, circular, prokaryoticallyderived
genome of -150 kb. The genome equivalents of plastid organelles in higher plant cells are an
attractive target for genetic engineering as high protein expression levels are readily obtained due to
the high genome copy number per organelle. The resultant proteins are contained within the plastid
organelle and the corresponding transgenes are inherited, in most crop plants, uniparentally,
preventing pollen transmission of DNA.
Plastid transformation involves the uniform modification of all the plastid genome copies, a process
facilitated by homologous recombination and the non-Mendelian segregation of plastids upon cell
division. The plastid genomes are in a continuous state of inter- and intra-molecular exchange due to
their common genetic complement. This enables the site-specific integration of any piece of DNA
flanked by plastid targeting sequences, via homologous recombination. The attainment of
homoplasmy, where all genomes are transformed, requires the inclusion of a plastid-specific selectable
marker. Selective pressure favouring the propagation of the transformed genome copies, as well as the
random segregation of plastids upon cell division, make it feasible to acquire uniformity and hence
genetic stability. From this, a complete transplastomie line is obtained where all plastid genome
copies present are transgenic, having eliminated all wild-type genome copies.
The prokaryotic nature of the chloroplast genetic system enables expression of multiple proteins from
polycistronic mRNAs, allowing the introduction of entire operons in a single transformation.
Expression cassettes in vectors thus include single regulatory elements of plastid origin, and harbour
genes encoding selectable and screenable markers, as well as one or more genes of interest. Each
coding region is preceded by an appropriate translation control region to ensure efficient translation
from the polycistronic mRNA.
The function of a plastid transformation vector is to enable transfer and stable integration of foreign
genes into the chloroplast genomes of higher plants. The expression vector constructed in this
research is specific for the transformation of the grape chloroplast genome. Vitis vinifera L., from the
family, Vitaceae, is the choice species for the production of wine and therefore our target for plastid
transformation. All chloroplast derived regulatory elements and sequences included in the vector thus
originated from this species. / AFRIKAANSE OPSOMMING: Die genetiese inligting van plante word gevind in die kern, die mitochondria, en die plastiede. Die
DNA van plastiede bestaan uit veelvuldige kopieë van 'n ~ 150 kb dubbelstring, sirkulêre genoom van
prokariotiese oorsprong. Die genoomekwivalente van plastiede in hoër plante is 'n aantreklike teiken
vir genetiese manipulering, aangesien die hoë genoom kopiegetal per organel dit moontlik maak om
gereeld hoë vlakke van proteïenuitdrukking te verkry. Hierdie proteïene word tot die plastied beperk,
en die ooreenstemmende transgene word in die meeste plante sitoplasmies oorgeërf, sonder die
oordrag van DNA deur die stuifmeel.
Plastied transformasie behels die uniforme modifikasie van al die plastied genoomkopieë, 'n proses
wat deur homoloë rekombinasie en die nie-Mendeliese segregasie van plastiede tydens seldeling
gefasiliteer word. As gevolg van die gemeenskaplike genetiese komplement, vind aanhoudende interen
intra-molekulêre uitruiling van plastiedgenome plaas. Dit maak die setel-spesifieke integrasie, via
homoloë rekombinasie, van enige stuk DNA wat deur plastied teikenvolgordes begrens word,
moontlik. Vir die verkrying van homoplasmie, waar alle genome getransformeer is, word die
insluiting van 'n plastiedspesifieke selekteerbare merker benodig. Seleksiedruk wat die vermeerdering
van die getransformeerde genoomkopieë bevoordeel, en die lukrake segregasie van plastiede tydens
seldeling, maak dit moontlik om genetiese stabiliteit en uniformiteit van die genoom te verkry. Dit
kan op sy beurt tot die verkryging van 'n volledige transplastomiese lyn lei, waar alle aanwesige
plastiedgenome transgenies is, en wilde tipe genoomkopieë geëlimineer is.
Die prokariotiese aard van die chloroplas genetiese sisteem maak die uitdrukking van veelvuldige
proteïene vanaf polisistroniese mRNAs moontlik, wat die toevoeging van volledige operons in 'n
enkele transformasie toelaat. Uitdrukkingskassette in vektore bevat dus enkel regulatoriese elemente
van plastied oorsprong, gene wat kodeer vir selekteerbare en sifbare merkers, asook een of meer gene
van belang (teikengene). Voor elke koderingsstreek, is daar ook 'n toepaslike translasie beheerstreek
om doeltreffende translasie vanaf die polisistroniese mRNA te verseker.
Die funksie van 'n plastied transformasie vektor is om die oordrag en stabiele integrasie van transgene
in chloroplasgenome van hoër plante moontlik te maak. Die uitdrukkingsvektor wat in hierdie studie
gekonstrueer is, is spesifiek vir die transformasie van die druif chloroplasgenoom. Vitis vinifera L.,
van die familie Vitaceae, is die voorkeur species vir die produksie van wyn, en daarom die teiken vir
plastied transformasie. Alle chloroplast-afgeleide regulatoriese elemente en volgordes wat in hierdie
vektor ingesluit is, het huloorsprong vanaf VUis vinifera L.
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Recombinant expression of the pRb- and p53-interacting domains from the human RBBP6 protein for in vitro binding studiesNdabambi, Nonkululeko January 2004 (has links)
Magister Scientiae - MSc / This thesis describes the cloning and recombinant expression of domains from the human RBBP6 protein for future in vitro binding studies with pRb and p53. RBBP6 is a splicing-associated protein that is known to interact with both p53 and the Retinoblastoma gene product (pRb), and has recently been shown to be highly upregulated in oesophageal cancer. The pRb binding domain (RbBD) and the p53 binding domain (p53BD) were each expressed using the glutathione-S-transferase (GST) tag affinity system, and affinity purified using a glutathione-linked agarose column. Purified fusion proteins were cleaved to separate the target protein from GST using PreScission™ Protease, for which there is a recognition sequence located immediately upstream of the multiple cloning site on the pGEX-6P series of plasmids. The pRb binding and p53 binding domains were further purified using cation exchange chromatography. Mass spectrometry confirmed that the RbBD was expressed as a single species of the expected molecular weight. However preliminary NMR analysis suggested that the domain was not fully folded. A total yield of 8 mg of protein was achieved from 1l of culture, which make it feasible to express 15N and 12C labelled samples for NMR. The p53BD was found to be expressed at lower levels and subject to C-terminal degradation, which suggest that the C-terminus is unstructured most likely due to the presence of poly-lysine tail. Human pRb protein was also successfully expressed and purified using the GST affinity system. Human p53 protein was expressed but was found to be insoluble and attempts to purify it were not pursued. Attempts to confirm the interactions between human RBBP6 and p53 and pRb proteins are on-going but fall outside the scope of this thesis. Expression constructs for the RING and zinc finger domains from human RBBP6 were also cloned into the pGEX system for future structural studies using NMR. Both domains were found to be expressed as soluble fusion proteins in preliminary expression studies. / South Africa
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Construction of a Cloning Vector Based upon a Rhizobium Plasmid Origin of Replication and its Application to Genetic Engineering of Rhizobium StrainsJeong, Pyengsoo 05 1900 (has links)
Rhizobia are Gram-negative, rod-shaped, soil bacteria with the ability to fix atmospheric nitrogen into ammonia as symbiont bacteroids within nodules of leguminous plant roots. Here, resident Rhizobium plasmids were studied as possible sources of components for the construction of a cloning vector for Rhizobium species.
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Selectable markers for recombinant poxvirusBrand, Elizabeth Gertruida 20 July 2017 (has links)
No description available.
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