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  1. Article: PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates.

    Barnes, W M

    Proceedings of the National Academy of Sciences of the United States of America

    1994  Volume 91, Issue 6, Page(s) 2216–2220

    Abstract: A target length limitation to PCR amplification of DNA has been identified and addressed. Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased. These improvements were ... ...

    Abstract A target length limitation to PCR amplification of DNA has been identified and addressed. Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased. These improvements were achieved by the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified to high yields from 1 ng of lambda DNA template.
    MeSH term(s) Bacteriophage lambda/genetics ; Base Composition ; Base Sequence ; DNA Primers ; DNA, Viral/analysis ; DNA, Viral/chemistry ; Exonucleases/metabolism ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Mutation ; Particle Size ; Polymerase Chain Reaction/methods ; Templates, Genetic
    Chemical Substances DNA Primers ; DNA, Viral ; Exonucleases (EC 3.1.-)
    Language English
    Publishing date 1994-03-15
    Publishing country United States
    Document type Journal Article
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.91.6.2216
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: The fidelity of Taq polymerase catalyzing PCR is improved by an N-terminal deletion.

    Barnes, W M

    Gene

    1992  Volume 112, Issue 1, Page(s) 29–35

    Abstract: KlenTaq DNA polymerase is an N-terminally truncated Thermus aquaticus (Taq) DNA polymerase I. As expressed from a gene construct in Escherichia coli, translation initiates at Met236, bypassing the 5'----3' exonuclease domain of the DNA polymerase- ... ...

    Abstract KlenTaq DNA polymerase is an N-terminally truncated Thermus aquaticus (Taq) DNA polymerase I. As expressed from a gene construct in Escherichia coli, translation initiates at Met236, bypassing the 5'----3' exonuclease domain of the DNA polymerase-encoding gene. A sensitive forward mutation assay was used to measure the relative number of mutations introduced into the entire lacZ gene by the polymerase chain reaction (PCR) under various conditions which allow the amplification of such a large DNA span. Two selectable markers, one at each end of the test lacZ fragment, were employed to avoid the plating and scoring of PCR artefacts such as primer initiation in the midst of the lacZ gene, and cloning artefacts such as empty vector plasmid. The measured relative mutation rate was twofold lower for KlenTaq as compared to the full-length Taq DNA polymerase.
    MeSH term(s) Base Sequence ; Cloning, Molecular ; DNA, Recombinant/metabolism ; DNA-Directed DNA Polymerase/metabolism ; Lac Operon ; Molecular Sequence Data ; Mutation/genetics ; Polymerase Chain Reaction/methods ; Taq Polymerase ; Thermus/enzymology ; beta-Galactosidase/genetics
    Chemical Substances DNA, Recombinant ; Taq Polymerase (EC 2.7.7.-) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; beta-Galactosidase (EC 3.2.1.23)
    Language English
    Publishing date 1992-03-01
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 391792-7
    ISSN 1879-0038 ; 0378-1119
    ISSN (online) 1879-0038
    ISSN 0378-1119
    DOI 10.1016/0378-1119(92)90299-5
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Variable patterns of expression of luciferase in transgenic tobacco leaves.

    Barnes, W M

    Proceedings of the National Academy of Sciences of the United States of America

    1990  Volume 87, Issue 23, Page(s) 9183–9187

    Abstract: A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic ... ...

    Abstract A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic plants. A military-type starlight vision system was used to conveniently analyze the pattern of gene expression in transgenic tobacco plant leaves. Transgenic tobacco plants which expressed luciferase uniformly in all areas of the leaf, and assays for luciferin, demonstrated that luciferin rapidly penetrates all regions of a tobacco leaf in at least two dimensions. Depending on the test gene structure or, presumably, on the transferred DNA (T-DNA) insertional context, other transgenic plants were obtained that expressed luciferase with a wide range of nonuniform patterns from nominally the same cauliflower mosaic virus 35S promoter. For instance, the veins can be dark, while only the interveinal regions of the leaf lamina glow, or only the small capillary veins glow, or only the major veins glow. Local and/or systemic induction in response to wounding was also demonstrated.
    MeSH term(s) Animals ; Coleoptera/enzymology ; Gene Expression ; Genetic Techniques ; Luciferases/genetics ; Luciferases/metabolism ; Plants/enzymology ; Plants/genetics ; Plants, Toxic ; Plasmids ; Promoter Regions, Genetic ; Rhizobium/genetics ; Nicotiana/enzymology ; Nicotiana/genetics
    Chemical Substances Luciferases (EC 1.13.12.-)
    Language English
    Publishing date 1990-12
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.87.23.9183
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Sequencing DNA with dideoxyribonucleotides as chain terminators: hints and strategies for big projects.

    Barnes, W M

    Methods in enzymology

    1987  Volume 152, Page(s) 538–556

    MeSH term(s) Base Sequence ; DNA/genetics ; DNA Nucleotidylexotransferase ; DNA-Directed DNA Polymerase ; Deoxyribonucleotides ; Exodeoxyribonucleases ; Indicators and Reagents ; Methods ; T-Phages/enzymology
    Chemical Substances Deoxyribonucleotides ; Indicators and Reagents ; DNA (9007-49-2) ; DNA Nucleotidylexotransferase (EC 2.7.7.31) ; DNA-Directed DNA Polymerase (EC 2.7.7.7) ; Exodeoxyribonucleases (EC 3.1.-) ; exodeoxyribonuclease III (EC 3.1.11.2)
    Language English
    Publishing date 1987
    Publishing country United States
    Document type Journal Article
    ISSN 0076-6879
    ISSN 0076-6879
    DOI 10.1016/0076-6879(87)52060-2
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  5. Article: Variable patterns of expression of luciferase in transgenic tobacco leaves

    Barnes, W.M

    Proceedings of the National Academy of Sciences of the United States of America. Dec 1990. v. 87 (23)

    1990  

    Abstract: A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic ... ...

    Abstract A carboxyl-terminally modified firefly luciferase, encoded as a gene fusion to the neomycin phosphotransferase gene (which confers kanamycin resistance), was found to be enzymatically active for both enzymes when expressed in bacteria and in transgenic plants. A military-type starlight vision system was used to conveniently analyze the pattern of gene expression in transgenic tobacco plant leaves. Transgenic tobacco plants which expressed luciferase uniformly in all areas of the leaf, and assays for luciferin, demonstrated that luciferin rapidly penetrates all regions of a tobacco leaf in at least two dimensions. Depending on the test gene structure or, presumably, on the transferred DNA (T-DNA) insertional context, other transgenic plants were obtained that expressed luciferase with a wide range of nonuniform patterns from nominally the same cauliflower mosaic virus 35S promoter. For instance, the veins can be dark, while only the interveinal regions of the leaf lamina glow, or only the small capillary veins glow, or only the major veins glow. Local and/or systemic induction in response to wounding was also demonstrated.
    Keywords Nicotiana tabacum ; leaves ; luciferase ; gene expression ; Agrobacterium tumefaciens ; Cauliflower mosaic virus ; disease resistance ; enzyme inhibitors ; Solanum lycopersicum var. lycopersicum ; proteinases ; genetically modified organisms ; gene fusion
    Language English
    Dates of publication 1990-12
    Size p. 9183-9187., ill.
    Document type Article
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    Database NAL-Catalogue (AGRICOLA)

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  6. Article: Cloning and restriction map of the first part of the histidine operon of Salmonella typhimurium.

    Barnes, W M

    Journal of bacteriology

    1981  Volume 147, Issue 1, Page(s) 124–134

    Abstract: The first part of the histidine operon of Salmonella typhimurium, hisGpeaGD, has been cloned onto the vector plasmid mini-ColE1 (pVH51). The resulting plasmid, pWB91, has a single EcoRI site and is 11,500 base pairs in size. The HindII restriction map ... ...

    Abstract The first part of the histidine operon of Salmonella typhimurium, hisGpeaGD, has been cloned onto the vector plasmid mini-ColE1 (pVH51). The resulting plasmid, pWB91, has a single EcoRI site and is 11,500 base pairs in size. The HindII restriction map was determined by the method of two-dimensional cross-annealing between a partial digest pattern and a complete digest pattern. The restriction fragment containing the genetic control region was identified with the aid of the small (35-base pair) internal deletion 01242 and the observation that heteroduplexed restriction fragments containing this deletion have markedly reduced mobility on polyacrylamide gels. The genetic control region was then mapped in more detail with other restriction enzymes. The genetic orientation of the restriction map was determined with the aid of several deletions of integral HindII fragments generated in vitro.
    MeSH term(s) Cloning, Molecular ; DNA Restriction Enzymes ; Deoxyribonucleases, Type II Site-Specific ; Genes, Regulator ; Histidine/biosynthesis ; Nucleic Acid Heteroduplexes ; Operon ; Plasmids ; Salmonella typhimurium/genetics
    Chemical Substances Nucleic Acid Heteroduplexes ; Histidine (4QD397987E) ; DNA Restriction Enzymes (EC 3.1.21.-) ; Deoxyribonucleases, Type II Site-Specific (EC 3.1.21.4) ; GTYRAC-specific type II deoxyribonucleases (EC 3.1.21.4)
    Language English
    Publishing date 1981-07
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2968-3
    ISSN 1098-5530 ; 0021-9193
    ISSN (online) 1098-5530
    ISSN 0021-9193
    DOI 10.1128/jb.147.1.124-134.1981
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  7. Article: Construction of an M13 histidine-transducing phage: a single-stranded cloning vehicle with one EcoRI site.

    Barnes, W M

    Gene

    1979  Volume 5, Issue 2, Page(s) 127–139

    Abstract: In order to create a ready source of single-stranded DNA for DNA sequence determination by the dideoxy chain-termination method, the promoter-proximal part of the histidine operon, the hisOGD region of Salmonella typhimurium, was cloned onto the single- ... ...

    Abstract In order to create a ready source of single-stranded DNA for DNA sequence determination by the dideoxy chain-termination method, the promoter-proximal part of the histidine operon, the hisOGD region of Salmonella typhimurium, was cloned onto the single-stranded phage M13. Both orientations of the his DNA were cloned to supply DNA template for sequencing of each strand. Insertion was achieved at an HaeIII site in the intergenic region (IR) of M13, and a single EcoRI site was purposely regenerated at one boundary of the his DNA insert. Infected colonies, not plaques, were selected using the hisD gene as a selective marker. The single RI site and the hisD marker for auxotrophic selection represent improvements on the wild type M13 as a single-stranded vector for cloning other DNA.
    MeSH term(s) Base Sequence ; DNA Restriction Enzymes/metabolism ; DNA, Recombinant ; DNA, Single-Stranded/genetics ; Histidine/genetics ; Nucleic Acid Hybridization ; Operon ; Salmonella typhimurium/genetics ; Transduction, Genetic
    Chemical Substances DNA, Recombinant ; DNA, Single-Stranded ; Histidine (4QD397987E) ; DNA Restriction Enzymes (EC 3.1.21.-)
    Language English
    Publishing date 1979-02
    Publishing country Netherlands
    Document type Journal Article ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 391792-7
    ISSN 1879-0038 ; 0378-1119
    ISSN (online) 1879-0038
    ISSN 0378-1119
    DOI 10.1016/0378-1119(79)90098-2
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Book: BASIC Surveying

    Barnes, W. M

    1988  

    Author's details W. M. Barnes
    Size 196 S
    Publisher Butterworths
    Publishing place London
    Document type Book
    ISBN 040801248X ; 9780408012485
    Database Library catalogue of the German National Library of Science and Technology (TIB), Hannover

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  9. Article: DNA sequencing by partial ribosubstitution.

    Barnes, W M

    Journal of molecular biology

    1978  Volume 119, Issue 1, Page(s) 83–99

    MeSH term(s) Base Sequence ; DNA/analysis ; DNA Polymerase I/metabolism ; Methods ; Ribonucleotides/analysis
    Chemical Substances Ribonucleotides ; DNA (9007-49-2) ; DNA Polymerase I (EC 2.7.7.-)
    Language English
    Publishing date 1978-02-15
    Publishing country England
    Document type Journal Article ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 80229-3
    ISSN 1089-8638 ; 0022-2836
    ISSN (online) 1089-8638
    ISSN 0022-2836
    DOI 10.1016/0022-2836(78)90271-1
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    Zs.A 867: Show issues Location:
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  10. Article: DNA sequence from the histidine operon control region: seven histidine codons in a row.

    Barnes, W M

    Proceedings of the National Academy of Sciences of the United States of America

    1978  Volume 75, Issue 9, Page(s) 4281–4285

    Abstract: The DNA sequence of 250 base pairs preceding the first structural gene of the histidine operon of Salmonella typhimurium was determined by the dideoxy chain-termination method. Single-stranded DNA template was provided by an M13-histidine transducing ... ...

    Abstract The DNA sequence of 250 base pairs preceding the first structural gene of the histidine operon of Salmonella typhimurium was determined by the dideoxy chain-termination method. Single-stranded DNA template was provided by an M13-histidine transducing phage constructed for the purpose by in vitro recombination. The termination site for the histidine leader RNA is identified by analogy with the trp operon leader termination sequence, and is 47 nucleotides before the start codon of the first structural gene G. Beginning 150 nucleotides before the end of the presumed leader RNA is a possible short protein-coding region with seven histidine codons in a row. It is proposed that the major mechanism of histodine operon control must involve a ribosome arrested at this run of histidine codons when histidine is limiting.
    MeSH term(s) Base Sequence ; Binding Sites ; Codon ; DNA, Bacterial/genetics ; Genes, Regulator ; Histidine/genetics ; Operon ; Ribosomes/metabolism ; Salmonella typhimurium/genetics
    Chemical Substances Codon ; DNA, Bacterial ; Histidine (4QD397987E)
    Language English
    Publishing date 1978-09
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.75.9.4281
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