Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 26 for “"T7 RNA polymerase"”.
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Transcription termination by bacteriophage T7 RNA polymerase at rho-independent terminators
… of transcription termination by bacteriophage RNA polymerases, templates encoding variants of the transcription-termination structure in the regulatory region of the threonine (thr) operon of Escherichia coli were used. The thr terminator comprises two distinct structural elements: a G+C-rich …
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Gene Expression Control for Synthetic Patterning of Bacterial Populations and Plants
… systems, I explore the use of bacteriophage T7 RNA polymerase as a gene expression engine to control synthetic patterning across populations of cells. In E. coli, I developed a ratiometric assay of bacteriophage T7 RNA polymerase activity, which I used to systematically characterize different …
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Physiological aspects underpinning recombinant protein production in Escherichia coli
… on the induction of very low levels of the T7 RNA polymerase in BL21* and thus the correspondingly slow synthesis of the target protein. Using this approach, growth and productivity of different types of correctly folded target proteins were sustained for at least 70 h. Secondly, mutant …
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Development of an inducible transcriptional control system in plasmodium falciparum with applications to targeted genome editing
… able to construct vectors to reconstitute the T7 RNA polymerase expression system in P. falciparum, functionally creating the first system for directed expression of non-coding RNA in the parasite. We were then able to use this expression system to adapt a clustered regularly interspaced short …
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Identification of Protein-Phosphate Contacts in the R17 Coat Protein Binding Site
Small RNAs of defined length and sequence can be synthesized enzymatically using T7 RNA polymerase and synthetic DNA templates which contain the T7 promoter. The templates are base paired from $-17$ to +1 and contain a long 5$\sp\prime$ overhang. The products of the transcription reactions have the …
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Specific Bacteriophage Coat Protein-Rna Interactions
… operator variants have been synthesized by using RNA ligase and transcription of synthetic DNA by T7 RNA polymerase in order to examine the roles of the helical region and the bulged A residue in the specific RNA-R17 coat protein interaction. The affinity between coat protein and each variant was …
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THE IDENTIFICATION OF NOVEL INORGANIC CARBON SENSITIVE ENZYMES
… active site with the adenylyl cyclases; the polymerase I family of prokaryotic DNA polymerases, the polymerase β family of DNA polymerases, and the guanylyl cyclases. Initial in vitro experiments performed on T7 RNA polymerase demonstrated a response to inorganic carbon, however, it was …
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Modifikuotų nukleotidų taikymas plataus masto nukleorūgščių analizei /
… that OTDDNs are substrates for a range of DNA polymerases of families A, B, X and RT. By means of in vitro evolution we have selected T7 RNA polymerase mutant with relaxed substrate discrimination that is able to synthesized chimeric nucleic acids labeled by OTDDNs. Moreover, DNA polymerases …
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The utility of modified nucleotides for high-throughput nucleic acid analysis /
… that OTDDNs are substrates for a range of DNA polymerases of families A, B, X and RT. By means of in vitro evolution we have selected T7 RNA polymerase mutant with relaxed substrate discrimination that is able to synthesized chimeric nucleic acids labeled by OTDDNs. Moreover, DNA polymerases …
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CRISPR-based Control of Biosynthetic Pathways and Genetic Biosensors, Informed by a Quantitative Investigation of Metabolic Burden
… with CRISPR transcriptional activation and T7 RNA polymerase in E. coli, finding that the overhead cost of CRISPR-based control is relatively high, but that expansion of the circuit to several control nodes that share the overhead cost is an inexpensive addition. We next analyze a set of …
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Identification and molecular characterisation of a putative gene for cell detaching factor from Trichomonas vaginalis.
… The ORF was transcribed in vitro using T7 RNA polymerase and translated in rabbit reticulocyte lysate in the presence of 35S-methionine. Several high molecular weight proteins (>200, 165, 150, 130, 118, 80, 47 kDa) were produced. Two polypeptides of 205 and 220 kDa were …
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Small Angle X-ray Scattering study of conformational changes in steroid responsive activator (SRA) RNA upon binding with suppressor protein SHARP
… The steroid receptor activator (SRA) RNA is a large RNA transcript indicated to be an epigenetic regulatory component in the steroid receptor coactivator-1 (SRC-1) acetyltransferase complex. It is<strong> </strong>needed for a response to steroid hormones by tumor cells. However, this …
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RNA tools for optimization of multi-protein genetic systems
… present an approach that leverages transacting RNA regulators to explore large gene expression spaces without de novo library construction. First, we developed six sRNAs whose strengths have been optimized against a set of 15nt "target" sequences that can be inserted upstream of a …
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Regulatory systems for the robust control of engineered genetic programs
… we build a 'resource allocator' by fragmenting T7 RNA polymerase variants into a conserved fragment and a set of variable fragments. The resource allocator limits the total number of polymerases that can be active in a genetic program, with the aim of protecting the host from being overburdened. …
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Dynamic Regulation of Metabolic Flux Using Orthogonal Quorum Sensing
… biology was further illustrated by using T7 RNA polymerase to amplify the expression output of an Agr-controlled circuit. Overall, this work developed and applied QS-based regulation systems to improve microbial production of value-added chemicals in E. coli.
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New strategies for in vivo continuous directed evolution
… fusions of various DNA damaging enzymes and the T7 RNA polymerase, achieved high mutation rates without the usual toxicity associated with off-target mutagenesis. We expect this mutagenesis technique to be applicable across a wide variety of organisms and particularly useful for viral-based …
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Reverse genetics system and fucntion of NSM protein of Rift Valley fever virus (family Bunyaviridae, genus phlebovirus)
… human vaccines are available. I established a T7 RNA polymerase-based reverse genetics system to rescue infectious clones of RVFV MP-12 strain entirely from cDNA, the first for any phleboviruses. Using this system, RVFV carrying substitution mutations of the M gene preglycoprotein region …
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Bistability, Synthetic Biology, and Antibiotic Treatment
… point by a simple circuit consisting of mutant T7 RNA polymerase (T7 RNAP*) that activates its own expression in bacterium Escherichia coli. Although activation by the T7 RNAP* is noncooperative, the circuit caused bistable gene expression. This counterintuitive observation can be explained by …
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Entwicklung neuartiger bakterieller Vektoren zur Übertragung von zellassoziierten Antigenen, DNA und RNA auf der Basis virulenzattenuierter L. monocytogenes
… Herkunft über das LPXTG-Ankermotiv von Internalin A. Die so erzielte starke Expression in der Zellwand setzte aber sowohl die Fitness der Bakterien als auch deren Invasivität in vitro deutlich herab und verhinderte damit eine effektive MHC-I-Präsentation des verwendeten Modellantigens. …
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