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

研究成果: Chapter in Book/Report/Conference proceedingChapter

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.

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

All Science Journal Classification (ASJC) codes

  • 免疫学および微生物学(全般)

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