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Analysis and optimization of recombinant DNA joining reactions

Identifieur interne : 004E10 ( Main/Exploration ); précédent : 004E09; suivant : 004E11

Analysis and optimization of recombinant DNA joining reactions

Auteurs : Randy J. Legerski [États-Unis] ; Donald L. Robberson [États-Unis]

Source :

Journal of Molecular Biology [ 0022-2836 ] ; 1985.

RBID : ISTEX:2CAB1FD8D9F9922E80247480BFFAF299AD5A37FB

English descriptors

Teeft :
- Appropriate antibiotic, Bacteriophage lambda, Bumhi site, Circular duplex, Circular plasmid vectors, Circular plasmids, Circular vectors, Clone, Cohesive ends, Collins hohn, Concatameric structures, Cosmid, Cyclic, Cyclic population, Cyclic structures, Different sizes, Different termini, Distribution functions, Dna, Duplex, Electron micrograph, Electron microscopy, Electrophoresis patterns, Endonuclease, Final results, Fragment, Fragment sizes, Fragments range, Genomic libraries, Hblol cells, Higher concentrations, Higher oligomers, Hohn, Hohn murray, Homopolymer system, Jacobson, Jacobson stockmayer, John wilson, Lambda, Lambda arms, Lambda recombinants, Legerski, Ligase, Linear molecules, Linear population, Lobban kaiser, Main text, Maximum percentage, Mole, Mole fraction, Molecular distributions, Molecular species, Molecular weight, Molecular weights, Molecule, Monomer, Nucleic acid, Oligomers, Optimal concentration, Optimal concentrations, Optimization, Packaging system, Plasmid, Polynucleotide ligase, Reasonable agreement, Recombinant, Recombinant dnas, Recombinant species, Restriction endonucleases, Restriction enzyme fragments, Restriction enzymes, Right arms, Robberson, Single restriction enzyme cleavage system, Statistical segment length, Stockmayer, Terminus, Theoretical curves, Total concentration, Total extent, Typical plot, Unique bamhi, Various sizes, Vector, Vector arms, Vector concentration, Vector fragment, Vector fragments, Vector molecule, Vector molecules, Vector size, Volume parameter, Wang, Wang davidson, Weight fraction.

Abstract

Abstract: The statistical segment length of duplex DNA was determined in phage T4 ligase (poly(deoxyribonucleotide): poly(deoxyribonucleotide) ligase (AMP forming), EC 6.5.1.1) buffer (50 mm-Tris · HCl (pH 7.8), 20 mm-dithiothreitol, 10 mm-MgCl2, 1 mm-ATP) at 12 °C to be 1030(± 116) Å. This result was obtained by electron microscopic examination of the molecular distributions generated by T4 ligase-mediated joining of EcoRI-cleaved pBR322 DNA. This value of the statistical segment length was utilized in an extension of the Jacobson-Stockmayer theory on the probability of intramolecular cyclization in order to optimize DNA joining reactions that are of great utility in recombinant DNA experiments. Five cloning systems were analyzed: circular plasmid vectors that had been linearized with one or two restriction endonucleases, circular plasmids that had been tailed with deoxyhomopolymers before joining, lambda-type cloning vectors and cosmids. The results are tabulated for convenient use in molecular cloning experiments.

Url:

https://api.istex.fr/ark:/67375/6H6-KPMPMV7Q-3/fulltext.pdf

DOI: 10.1016/0022-2836(85)90093-2

Affiliations:

États-Unis

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Le document en format XML

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<term>Circular plasmid vectors</term>
<term>Circular plasmids</term>
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<term>Cohesive ends</term>
<term>Collins hohn</term>
<term>Concatameric structures</term>
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<term>Cyclic population</term>
<term>Cyclic structures</term>
<term>Different sizes</term>
<term>Different termini</term>
<term>Distribution functions</term>
<term>Dna</term>
<term>Duplex</term>
<term>Electron micrograph</term>
<term>Electron microscopy</term>
<term>Electrophoresis patterns</term>
<term>Endonuclease</term>
<term>Final results</term>
<term>Fragment</term>
<term>Fragment sizes</term>
<term>Fragments range</term>
<term>Genomic libraries</term>
<term>Hblol cells</term>
<term>Higher concentrations</term>
<term>Higher oligomers</term>
<term>Hohn</term>
<term>Hohn murray</term>
<term>Homopolymer system</term>
<term>Jacobson</term>
<term>Jacobson stockmayer</term>
<term>John wilson</term>
<term>Lambda</term>
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<term>Lambda recombinants</term>
<term>Legerski</term>
<term>Ligase</term>
<term>Linear molecules</term>
<term>Linear population</term>
<term>Lobban kaiser</term>
<term>Main text</term>
<term>Maximum percentage</term>
<term>Mole</term>
<term>Mole fraction</term>
<term>Molecular distributions</term>
<term>Molecular species</term>
<term>Molecular weight</term>
<term>Molecular weights</term>
<term>Molecule</term>
<term>Monomer</term>
<term>Nucleic acid</term>
<term>Oligomers</term>
<term>Optimal concentration</term>
<term>Optimal concentrations</term>
<term>Optimization</term>
<term>Packaging system</term>
<term>Plasmid</term>
<term>Polynucleotide ligase</term>
<term>Reasonable agreement</term>
<term>Recombinant</term>
<term>Recombinant dnas</term>
<term>Recombinant species</term>
<term>Restriction endonucleases</term>
<term>Restriction enzyme fragments</term>
<term>Restriction enzymes</term>
<term>Right arms</term>
<term>Robberson</term>
<term>Single restriction enzyme cleavage system</term>
<term>Statistical segment length</term>
<term>Stockmayer</term>
<term>Terminus</term>
<term>Theoretical curves</term>
<term>Total concentration</term>
<term>Total extent</term>
<term>Typical plot</term>
<term>Unique bamhi</term>
<term>Various sizes</term>
<term>Vector</term>
<term>Vector arms</term>
<term>Vector concentration</term>
<term>Vector fragment</term>
<term>Vector fragments</term>
<term>Vector molecule</term>
<term>Vector molecules</term>
<term>Vector size</term>
<term>Volume parameter</term>
<term>Wang</term>
<term>Wang davidson</term>
<term>Weight fraction</term>
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<front><div type="abstract" xml:lang="en">Abstract: The statistical segment length of duplex DNA was determined in phage T4 ligase (poly(deoxyribonucleotide): poly(deoxyribonucleotide) ligase (AMP forming), EC 6.5.1.1) buffer (50 mm-Tris · HCl (pH 7.8), 20 mm-dithiothreitol, 10 mm-MgCl2, 1 mm-ATP) at 12 °C to be 1030(± 116) Å. This result was obtained by electron microscopic examination of the molecular distributions generated by T4 ligase-mediated joining of EcoRI-cleaved pBR322 DNA. This value of the statistical segment length was utilized in an extension of the Jacobson-Stockmayer theory on the probability of intramolecular cyclization in order to optimize DNA joining reactions that are of great utility in recombinant DNA experiments. Five cloning systems were analyzed: circular plasmid vectors that had been linearized with one or two restriction endonucleases, circular plasmids that had been tailed with deoxyhomopolymers before joining, lambda-type cloning vectors and cosmids. The results are tabulated for convenient use in molecular cloning experiments.</div>
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Analysis and optimization of recombinant DNA joining reactions

Analysis and optimization of recombinant DNA joining reactions

Source :

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri