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2013
Site-directed mutagenesis is a PCR-based method to mutate specified nucleotides of a sequence within a plasmid vector. This technique allows one to study the relative importance of a particular amino acid for protein structure and function. Typical mutations are designed to disrupt or map protein-protein interactions, mimic or block posttranslational ...
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Site-directed mutagenesis is a PCR-based method to mutate specified nucleotides of a sequence within a plasmid vector. This technique allows one to study the relative importance of a particular amino acid for protein structure and function. Typical mutations are designed to disrupt or map protein-protein interactions, mimic or block posttranslational ...
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1983
In the preceding chapters dealing with modifications of nucleic acids it is assumed that, in general, these chemical reactions are randomly located along the genome. Exceptions occur when protein-nucleic interactions or nucleic acid secondary structure playa role.
B. Singer, D. Grunberger
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In the preceding chapters dealing with modifications of nucleic acids it is assumed that, in general, these chemical reactions are randomly located along the genome. Exceptions occur when protein-nucleic interactions or nucleic acid secondary structure playa role.
B. Singer, D. Grunberger
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Inverse PCR for Site-Directed Mutagenesis
2023Inverse PCR is a powerful tool for the rapid introduction of desired mutations at desired positions in a circular double-stranded DNA sequence. In this technique, custom-designed mutant primers oriented in the inverse direction are used to amplify the entire circular template with incorporation of the required mutation(s).
Diogo, Silva +4 more
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Site-Directed Mutagenesis in the Mouse
1993Publisher Summary This chapter presents site-directed mutagenesis in the mouse. The correlation of genotype and phenotype is a central theme in the analysis of development. In the mouse, this type of reverse genetics is constrained not only by the size of the murine genome, which makes positional cloning difficult, but also by the limited ...
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Site-Directed Mutagenesis of Bifidobacterium Strains
2021At present, only a limited number of Bifidobacterium species are amenable to genetic manipulation using mutagenesis. This lack of genetic accessibility among the majority of bifidobacterial strains represents a significant roadblock for the study of gene function and expression in these potential probiotics.
Kieran, James, Douwe, van Sinderen
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Philosophical Transactions of the Royal Society of London. Series A, Mathematical and Physical Sciences, 1982
Abstract The development of genetic engineering, whereby a specific gene or cDNA (c is copy or complementary) can be isolated as part of a minichromosome that can replicate and be expressed (by transcription and translation) in a living cell, has made possible in vitro techniques for micromanipulation (i.e.
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Abstract The development of genetic engineering, whereby a specific gene or cDNA (c is copy or complementary) can be isolated as part of a minichromosome that can replicate and be expressed (by transcription and translation) in a living cell, has made possible in vitro techniques for micromanipulation (i.e.
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Trends in Plant Science, 2000
A system has been developed for site-directed mutagenesis in plants. It has been dubbed the pinpoint technique because it takes advantage of the plant’s DNA repair system and makes use of specifically designed hairpin-shaped molecules of DNA and RNA.
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A system has been developed for site-directed mutagenesis in plants. It has been dubbed the pinpoint technique because it takes advantage of the plant’s DNA repair system and makes use of specifically designed hairpin-shaped molecules of DNA and RNA.
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PCR-Mediated Site-Directed Mutagenesis
Cold Spring Harbor Protocols, 2013Unlike traditional site-directed mutagenesis, this protocol requires only a single PCR step using full plasmid amplification to generate point mutants. The method can introduce small mutations into promoter sites and is even better suited for introducing single or double mutations into proteins.
Michael F, Carey +2 more
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Activation of aspartase by site-directed mutagenesis
Biochemical and Biophysical Research Communications, 1991To elucidate the role of sulfhydryl groups in the enzymatic reaction of the aspartase from Escherichia coli, we used site-directed mutagenesis which showed that the enzyme was activated by replacement of Cys-430 with a tryptophan. This mutation produced functional alterations without appreciable structural change: The kcat values became 3-fold at pH 6 ...
S, Murase +5 more
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High-Throughput Site-Directed Mutagenesis
2019Protein engineering has an array of uses: whether you are studying a disease mutation, removing undesirable sequences, adding stabilizing mutations for structural purposes, or simply dissecting protein function. Protein engineering is almost exclusively performed using site-directed mutagenesis (SDM) as this provides targeted modification of specific ...
Claire, Strain-Damerell +1 more
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