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Segment polarity genes in Drosophila developmentForbes, Alexandria J. January 1992 (has links)
No description available.
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Development of an in vivo DNA Cloning Procedure for Bacteria / Development of an in vivo DNA Cloning ProcedureChain, Patrick 12 1900 (has links)
In this thesis, we describe the development of a method to delete and to clone specific large regions from the 1700 kilobase pExo megaplasmid of Rhizobium meliloti. In principal, the region to be cloned is flanked by FRT sites, which direct site specific recombination by the Flp recombinase. Targeting constructs were designed to include part of the IS50 from Tn5, the FRT site, an origin of transfer (oriT), and a replication origin from RK2 or the F plasmid. These constructs were directed to known Tn5-derivative insertion sites in the pExo megaplasmid. A plasmid which expresses the Flp recombinase constitutively in R. meliloti was made, and the transfer of this plasmid to FRT-flanked megaplasmid regions was shown to result in the deletion of the intervening DNA. We demonstrated that the pExo megaplasmid DNA regions could be captured in Escherichia coli, however in this case, the megaplasmid excision event appears to be directed by the oriT sites rather than the FRT sites. We present strong evidence that a specific 50 kb region contains the oriV of the pExo megaplasmid; R. meliloti strains deleted for this region could not be isolated, and this region was found to replicate autonomously in Agrobacterium tumefaciens. Preliminary sequence analysis has revealed strong homology within this region to genes encoding the RepABC replication proteins of several Rhizobium and Agrobacterium plasmids. / Thesis / Master of Science (MS)
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Diagnostically significant antigens of Treponema pallidum subsp. pallidum : identification, serological efficacy, and characterisation of the major antigenic determinantsGrace, Christopher January 2000 (has links)
No description available.
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Molecular studies on the control of the expression of the NPY-1 receptor geneBournat, Juan Carlos January 1997 (has links)
No description available.
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DNA restriction fragment lenth polymorphisms in the identification of clonal variants of eucalyptus.Coulson, Mornay. January 1993 (has links)
The technique of restriction fragment length polymorphism (RFLP) analysis, of
chloroplastic and genomic DNA, was investigated as a means of identifying eucalypt species and cultivars which are morphologically indistinguishable from one another. In order to resolve chloroplast DNA (cpDNA) RFLPs, a method was developed to extract high yields of intact chloroplasts from Eucalyptus grandis S/N M6. Starch contamination was reduced by incubation of saplings in the dark for 48 h prior to extraction and watering with a solution containing 370 mM Na-phosphate and 296 mM KN03. Optimal chloroplast yields (25 ug chlorophyll/g fresh mass) were obtained by chopping leaf material, using a vertical homogenizer, in a buffer
containing 350 mM sorbitol, 50 mM tris-HCL and 5 mM EDTA, 0.1 % (w/v) bovine
serum albumin, 0.15 % (w/v) 2-mercaptoethanol, 2 mM L-ascorbic acid and 1 mM
MgCI2 followed by washing of leaf pieces in a buffer containing only sorbitol, tris-HCL and EDTA. When these chloroplasts were used in an "in-organelle" DNA
digestion procedure, polymorphisms were observed between the cpDNA profiles
resolved for E. grandis S/N M6 and that of an outgroup species (spinach). However,
the developed chloroplast extraction technique could not be used to obtain
chloroplasts from various other eucalypt species, probably as a result of variability in the material at an ultrastructural or biochemical level. For the analysis of genomic DNA RFLPs, a DNA extraction procedure was optimized for use with various eucalypt species and cultivars. This included the development of a purifcation technique during which DNA was ammonium acetate-ethanol
precipitated and subjected to mini-dialysis. Following Dra I restriction of DNA, the extract was electrophoresed and Southern blotted onto both nylon and nitrocellulose membranes. These were probed with a Hind-III restricted sample of
the multilocus plasmid probe pV47-2. This probe was labelled using 32p as well as
a non-radioactive labelling substance digoxygenin (DIG). Hybridization conditions,
including the composition of the hybridization buffer, were optimized for use with
these labels, and DNA RFLPs (fingerprints) were resolved for the eucalypt species
E. grandis and E. macarthurii and cultivars of E. grandis (S/N M6, TAG 5 and TAG
14). An average of 8.5 bands were detected with 32p and 5.0 fragments with DIG.
All the species and cultivars fingerprinted with the 32P-label could be distinguished
from one another. However, as a result of the reduced sensitivity of the DIG
system, two of the E. grandis cultivars, S/N M6 and TAG 5, could not be
differentiated. It is concluded that the latter system would be most suitable for
incorporation into a routine eucalypt screening programme, although it is suggested
that the colourimetric detection assay, used in this study to resolve DNA bands, be
replaced by a more sensitive one. / Thesis (M.Sc)-University of Natal, Durban, 1993.
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Expressao de endostatina em fibroblastos murinos para tratamento de tumores solidosTORNIERI, PAULA H. 09 October 2014 (has links)
Made available in DSpace on 2014-10-09T12:48:41Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:01:04Z (GMT). No. of bitstreams: 1
09151.pdf: 3898995 bytes, checksum: c6107a524057a95f300eee8a635c6f1e (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP
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Obtenção de um modelo homólogo de terapia gênica mediante administração direta de um plasmídeo com o gene do hormônio de crescimento murino em camundongos anões imunocompetentes / An homologous model of gene therapy by in vivo administration of a plasmid containing the mouse growth hormone gene in immunocompetent dwarf miceCECCHI, CLAUDIA R. 09 October 2014 (has links)
Made available in DSpace on 2014-10-09T12:35:39Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:56Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP
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Expressao de endostatina em fibroblastos murinos para tratamento de tumores solidosTORNIERI, PAULA H. 09 October 2014 (has links)
Made available in DSpace on 2014-10-09T12:48:41Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:01:04Z (GMT). No. of bitstreams: 1
09151.pdf: 3898995 bytes, checksum: c6107a524057a95f300eee8a635c6f1e (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP
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Obtenção de um modelo homólogo de terapia gênica mediante administração direta de um plasmídeo com o gene do hormônio de crescimento murino em camundongos anões imunocompetentes / An homologous model of gene therapy by in vivo administration of a plasmid containing the mouse growth hormone gene in immunocompetent dwarf miceCECCHI, CLAUDIA R. 09 October 2014 (has links)
Made available in DSpace on 2014-10-09T12:35:39Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:56Z (GMT). No. of bitstreams: 0 / Níveis sustentáveis de hormônio de crescimento humano (hGH) circulante e aumento de peso altamente significativo, avaliados também em comparação a repetidas injeções de hormônio, foram observados em trabalhos anteriores, baseados na eletrotransferência de DNA plasmidial no músculo de camundongos anões imunodeficientes (lit/scid). No presente trabalho, um modelo animal homólogo de terapia gênica para GH foi estudado mediante clonagem da sequência genômica do DNA de GH de camundongo (mGH-gDNA), a qual substituiu o hGH-gDNA no vetor que havia sido utilizado em camundongos anões imunodeficientes. O novo vetor, agora nomeado UBI-mGH-gDNA, foi utilizado em camundongos anões imunocompetentes (lit/lit). Foi primeiramente realizado um teste in vitro, transfectando-se células humanas HEK 293 com este plasmídeo e obtendo-se uma expressão de 3,0 μg mGH/106 células/dia, contra 3,7 μg mGH/106 células/dia, para o UBI-hGH-gDNA. Estes dois plasmídeos foram então injetados (50 μg/animal) no músculo quadríceps de camundongos, seguido de eletroporação, realizando um ensaio de 94 dias. Enquanto após 15 dias, as inclinações das curvas de variação de peso relacionadas ao mGH, hGH e salina foram 0,130, 0,112 e 0,027 g/camundongo/dia, respectivamente, após 94 dias, as inclinações correspondentes foram 0,041, 0,028 e 0,033 g/camundongo/dia. As análises estatísticas mostraram que após 15 dias, as inclinações das duas curvas com o GH foram significativamente maiores que a inclinação do controle (P<0,001), enquanto que após 94 dias, somente a inclinação da curva do mGH foi maior que a do controle (P<0,005). A porcentagem de aumento de peso nos animais tratados com o gene do mGH, após 94 dias, foi de 34,3%, enquanto que o comprimento nariz-cauda e o comprimento do fêmur, dois parâmetros que medem diretamente o crescimento longitudinal, foram de 9,5% e 26%, respectivamente, quando comparados aos valores iniciais. A interrupção do crescimento progressivo do grupo tratado com hGH não foi inesperada, considerando a óbvia reação imunogênica dos animais imunocompetentes contra o GH humano e não contra o de camundongo (título do anticorpo anti-hGH 1:100 a 1:3200). A inclinação altamente positiva do grupo controle, já observada em camundongos lit/lit mas não em lit/scid, é provavelmente devida ao ganho de peso natural desta linhagem, não suportada, contudo, por um proporcional crescimento longitudinal. Níveis circulatórios de mGH da ordem de 4 ng/mL foram detectados após 15 dias para o grupo tratado com o mGH, enquanto o grupo controle apresentou níveis em torno de 0,7 ng mGH/mL (P<0,001). Níveis circulatórios de mIGF-I foram também determinados nos dias 15, 45 e 94 nos animais tratados com mGH, sempre mostrando valores 1,5 - 3,0 vezes maiores que o grupo controle, e valores 1,2-1,6 vezes maiores que o grupo tratado com hGH. Este modelo de tratamento homólogo pode ser considerado uma primeira abordagem e um importante suporte para futuros ensaios pré-clínicos baseados na administração de DNA plasmidial para o tratamento da deficiência de GH humano. / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP
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Clonagem, caracterização e análise filogenética das subunidades alfa e beta do hormônio folículo estimulante de Pirarucu (Arapalma gigas) visando sua síntese em células CHO / Cloning, characterization and phylogenetic analysis of the alpha and beta subunits of the follicle stimulating hormone of Pirarucu (Arapalma gigas) in view of its synthesis in CHO cellsCARVALHO, ROBERTO F. de 10 November 2014 (has links)
Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2014-11-10T11:09:13Z
No. of bitstreams: 0 / Made available in DSpace on 2014-11-10T11:09:13Z (GMT). No. of bitstreams: 0 / Dissertação (Mestrado em Tecnologia Nuclear) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP
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