Application of environmental DNA resources to create useful DNA polymerases with different properties

研究成果: 著書/レポートタイプへの貢献

抄録

DNA polymerases use deoxynucleotide triphosphates to synthesize new DNA strands according to the template DNA during DNA replication and repair, and are essential to maintain genome integrity in DNA metabolism. In addition, these enzymes are widely used for genetic engineering techniques, including dideoxy-sequencing, PCR, DNA labeling, mutagenesis, and other in vitro gene manipulations. Thermostable DNA polymerases are especially useful for PCR and cycle-sequencing. We describe in this chapter a powerful strategy to use environmental DNA as a genetic resource to create useful DNA polymerases. The region corresponding to the active center of the DNA polymerizing reaction in the structural gene of well known DNA polymerases, such as Pfu DNA polymerase and Taq DNA polymerase, can be substituted with gene fragments amplified by PCR from DNAs within soil samples from various world-wide locations. The constructed chimeric pol genes can be expressed in E. coli, and the produced chimeric enzymes, possessing DNA polymerase activities with different properties, can be evaluated in terms of their processivity, fidelity, and efficiency of primer usage, to select valuable DNA polymerases for genetic engineering techniques.

元の言語英語
ホスト出版物のタイトルMicroorganisms in Sustainable Agriculture and Biotechnology
出版者Springer Netherlands
ページ663-678
ページ数16
9789400722149
ISBN(電子版)9789400722149
ISBN(印刷物)9400722133, 9789400722132
DOI
出版物ステータス出版済み - 10 1 2012

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DNA-Directed DNA Polymerase
DNA
Genetic Techniques
Genetic Engineering
Polymerase Chain Reaction
Taq Polymerase
pol Genes
Genes
Enzymes
DNA Replication
DNA Sequence Analysis
Mutagenesis
DNA Repair
Soil
Genome
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Immunology and Microbiology(all)

これを引用

Ishino, S., & Ishino, Y. (2012). Application of environmental DNA resources to create useful DNA polymerases with different properties. : Microorganisms in Sustainable Agriculture and Biotechnology (巻 9789400722149, pp. 663-678). Springer Netherlands. https://doi.org/10.1007/978-94-007-2214-9_29

Application of environmental DNA resources to create useful DNA polymerases with different properties. / Ishino, Sonoko; Ishino, Yoshizumi.

Microorganisms in Sustainable Agriculture and Biotechnology. 巻 9789400722149 Springer Netherlands, 2012. p. 663-678.

研究成果: 著書/レポートタイプへの貢献

Ishino, S & Ishino, Y 2012, Application of environmental DNA resources to create useful DNA polymerases with different properties. : Microorganisms in Sustainable Agriculture and Biotechnology. 巻. 9789400722149, Springer Netherlands, pp. 663-678. https://doi.org/10.1007/978-94-007-2214-9_29
Ishino S, Ishino Y. Application of environmental DNA resources to create useful DNA polymerases with different properties. : Microorganisms in Sustainable Agriculture and Biotechnology. 巻 9789400722149. Springer Netherlands. 2012. p. 663-678 https://doi.org/10.1007/978-94-007-2214-9_29
Ishino, Sonoko ; Ishino, Yoshizumi. / Application of environmental DNA resources to create useful DNA polymerases with different properties. Microorganisms in Sustainable Agriculture and Biotechnology. 巻 9789400722149 Springer Netherlands, 2012. pp. 663-678
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