An alternative method for the synthesis of tailor-made genes
Identifieur interne : 003F93 ( Main/Exploration ); précédent : 003F92; suivant : 003F94An alternative method for the synthesis of tailor-made genes
Auteurs : O. W. Liew [Nouvelle-Zélande] ; D. W. Bullock [Nouvelle-Zélande]Source :
- Plant Molecular Biology Reporter [ 0735-9640 ] ; 1996-03-01.
English descriptors
- KwdEn :
- Teeft :
- Alternative method, Amino acid, Amino acid sequence, Amplification, Base changes, Base substitutions, Clone, Cold spring harbor, Cold spring harbor laboratory, Crude mixture, Crude synthesis mixture, Deae membrane, Error rate, Exact fusion, Failure sequences, Gene synthesis, Oligonucleotide, Oligonucleotide synthesis, Polymerase, Polymerase chain reaction, Present work, Primer, Recognition sequence, Restriction cleavage, Restriction sites, Second round, Structural genes, Synthetic fragment, Synthetic oligonucleotide.
Abstract
Abstract: Oligonucleotide synthesis was coupled with amplification by the polymerase chain reaction to generate an exact translational fusion between a plant signal sequence and an animal structural gene. A synthetic 111-mer oligonucleotide representing less than two percent of the reaction products was successfully amplified by using short primers containing restriction sites designed for ease of cloning and providing in-frame fusion. The method overcomes the length-versus-yield dilemma in oligonucleotide synthesis, and is generally adaptable to the construction of a translationally competent coding sequence from any two DNA fragments.
Url:
DOI: 10.1007/BF02671899
Affiliations:
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W." last="Liew">O. W. Liew</name><affiliation wicri:level="1"><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Centre for Molecular Biology, Lincoln University, PO Box 84, Canterbury</wicri:regionArea><wicri:noRegion>Canterbury</wicri:noRegion></affiliation></author><author><name sortKey="Bullock, D W" sort="Bullock, D W" uniqKey="Bullock D" first="D. W." last="Bullock">D. W. Bullock</name><affiliation wicri:level="1"><country xml:lang="fr">Nouvelle-Zélande</country><wicri:regionArea>Centre for Molecular Biology, Lincoln University, PO Box 84, Canterbury</wicri:regionArea><wicri:noRegion>Canterbury</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Nouvelle-Zélande</country></affiliation></author></analytic><monogr></monogr><series><title level="j">Plant Molecular Biology Reporter</title><title level="j" type="abbrev">Plant Mol Biol Rep</title><idno type="ISSN">0735-9640</idno><idno type="eISSN">1572-9818</idno><imprint><publisher>Springer Netherlands</publisher><pubPlace>Dordrecht</pubPlace><date type="published" when="1996-03-01">1996-03-01</date><biblScope unit="volume">14</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="23">23</biblScope><biblScope unit="page" to="32">32</biblScope></imprint><idno type="ISSN">0735-9640</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0735-9640</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>gene fusions</term><term>oligonucleotide synthesis</term><term>polymerase chain reaction</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Alternative method</term><term>Amino acid</term><term>Amino acid sequence</term><term>Amplification</term><term>Base changes</term><term>Base substitutions</term><term>Clone</term><term>Cold spring harbor</term><term>Cold spring harbor laboratory</term><term>Crude mixture</term><term>Crude synthesis mixture</term><term>Deae membrane</term><term>Error rate</term><term>Exact fusion</term><term>Failure sequences</term><term>Gene synthesis</term><term>Oligonucleotide</term><term>Oligonucleotide synthesis</term><term>Polymerase</term><term>Polymerase chain reaction</term><term>Present work</term><term>Primer</term><term>Recognition sequence</term><term>Restriction cleavage</term><term>Restriction sites</term><term>Second round</term><term>Structural genes</term><term>Synthetic fragment</term><term>Synthetic oligonucleotide</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Oligonucleotide synthesis was coupled with amplification by the polymerase chain reaction to generate an exact translational fusion between a plant signal sequence and an animal structural gene. A synthetic 111-mer oligonucleotide representing less than two percent of the reaction products was successfully amplified by using short primers containing restriction sites designed for ease of cloning and providing in-frame fusion. The method overcomes the length-versus-yield dilemma in oligonucleotide synthesis, and is generally adaptable to the construction of a translationally competent coding sequence from any two DNA fragments.</div></front></TEI><affiliations><list><country><li>Nouvelle-Zélande</li></country></list><tree><country name="Nouvelle-Zélande"><noRegion><name sortKey="Liew, O W" sort="Liew, O W" uniqKey="Liew O" first="O. W." last="Liew">O. W. Liew</name></noRegion><name sortKey="Bullock, D W" sort="Bullock, D W" uniqKey="Bullock D" first="D. W." last="Bullock">D. W. Bullock</name><name sortKey="Bullock, D W" sort="Bullock, D W" uniqKey="Bullock D" first="D. W." last="Bullock">D. W. 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