Dna Shuffling
Mostrando 13-24 de 117 artigos, teses e dissertações.
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13. Expressão heteróloga, purificação e estudo de atividade de uma proteína inibidora de cisteínio protease da cana-de-açúcar e posterior evolução in vitro pela técnica de DNA Shuffling.
As plantas parecem ter desenvolvido ao longo da evolução mecanismos naturais de defesa contra fungos e insetos. Entre eles está o uso de inibidores de protease. Cistatinas são proteínas que inibem especificamente cisteíno proteases. Elas ocorrem naturalmente em várias espécies de vegetais e acredita-se que elas estejam envolvidas com mecanismos de de
Publicado em: 2004
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14. Construction of a mutant library of horseradish peroxidase gene by directed evolution and development of an in situ screening method
A process of directed evolution applied to obtain a library of mutants of horseradish peroxidase (HRP) enzyme is described. We have introduced slight variations into the original DNA shuffling protocol. A DNA template was prepared by PCR amplification and digested with Dnase I during 1 hour. Dnase I products were concentrated by precipitation with isopropano
Brazilian Journal of Chemical Engineering. Publicado em: 2003-03
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15. Transgênicos e evolução dirigida
Os eventos de mutação são responsáveis pela geração de variabilidade genética nas populações, permitindo a ação da seleção natural que favorece os tipos mais adaptados. A exploração dessa variabilidade, embora de forma empírica, teve início há cerca de dez mil anos, quando se deu a domesticação das primeiras culturas agrícolas. Com o adv
História, Ciências, Saúde-Manguinhos. Publicado em: 2000-10
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16. Computational and experimental analysis of DNA shuffling
We describe a computational model of DNA shuffling based on the thermodynamics and kinetics of this process. The model independently tracks a representative ensemble of DNA molecules and records their states at every stage of a shuffling reaction. These data can subsequently be analyzed to yield information on any relevant metric, including reassembly effici
The National Academy of Sciences.
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17. Optimization of DNA shuffling for high fidelity recombination.
A convenient 'DNA shuffling' protocol for random recombination of homologous genes in vitro with a very low rate of associated point mutagenesis (0.05%) is described. In addition, the mutagenesis rate can be controlled over a wide range by the inclusion of Mn2+or Mg2+during DNase I digestion, by choice of DNA polymerase used during gene reassembly as well as
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18. Random DNA fragmentation with endonuclease V: application to DNA shuffling
The enzyme endonuclease V nicks uracil-containing DNA at the second or third phosphodiester bond 3′ to uracil sites. I applied the enzyme to random fragmentation of DNA to revise the complex DNA shuffling protocol. The merit of using endonuclease V is that cleavage occurs at random sites and the length of the fragments can easily be adjusted by varying the
Oxford University Press.
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19. Directed evolution of a fucosidase from a galactosidase by DNA shuffling and screening
An efficient β-fucosidase was evolved by DNA shuffling from the Escherichia coli lacZ β-galactosidase. Seven rounds of DNA shuffling and colony screening on chromogenic fucose substrates were performed, using 10,000 colonies per round. Compared with native β-galactosidase, the evolved enzyme purified from cells from the final round showed a 1,000-fold inc
The National Academy of Sciences of the USA.
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20. Exon shuffling and the origin of the mitochondrial targeting function in plant cytochrome c1 precursor.
Since most of the examples of "exon shuffling" are between vertebrate genes, the view is often expressed that exon shuffling is limited to the evolutionarily recent lineage of vertebrates. Although exon shuffling in plants has been inferred from the analysis of intron phases of plant genes [Long, M., Rosenberg, C. & Gilbert, W. (1995) Proc. Natl. Acad. Sci.
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21. Creating multiple-crossover DNA libraries independent of sequence identity
We have developed, experimentally implemented, and modeled in silico a methodology named SCRATCHY that enables the combinatorial engineering of target proteins, independent of sequence identity. The approach combines two methods for recombining genes: incremental truncation for the creation of hybrid enzymes and DNA shuffling. First, incremental truncat
The National Academy of Sciences.
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22. Predicting crossover generation in DNA shuffling
We introduce a quantitative framework for assessing the generation of crossovers in DNA shuffling experiments. The approach uses free energy calculations and complete sequence information to model the annealing process. Statistics obtained for the annealing events then are combined with a reassembly algorithm to infer crossover allocation in the reassem
The National Academy of Sciences.
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23. Family Shuffling of Expandase Genes To Enhance Substrate Specificity for Penicillin G
Deacetoxycephalosporin C synthase (expandase) from Streptomyces clavuligerus, encoded by cefE, is an important industrial enzyme for the production of 7-aminodeacetoxycephalosporanic acid from penicillin G. To improve the substrate specificity for penicillin G, eight cefE-homologous genes were directly evolved by using the DNA shuffling technique. After the
American Society for Microbiology.
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24. Kinetic evidence that echinomycin migrates between potential DNA binding sites.
The hypothesis that echinomycin locates its preferred nucleotide sequences in DNA by a process of "shuffling" between potential binding sites has been tested. Immediately after reacting with calf thymus DNA the antibiotic is relatively weakly bound inasmuch as the complex dissociates quite rapidly when detergent is added. If the complex is allowed to equilib