Isothermal DNA amplification in vitro: the helicase-dependent amplification system.
Since the development of polymerase chain reaction, amplification of nucleic acids has emerged as an elemental tool for molecular biology, genomics, and biotechnology. Amplification methods often use temperature cycling to exponentially amplify nucleic acids; however, isothermal amplification methods have also been developed, which do not require heating the double-stranded nucleic acid to dissociate the synthesized products from templates. Among the several methods used for isothermal DNA amplification, the helicase-dependent amplification (HDA) is discussed in this review with an emphasis on the reconstituted DNA replication system. Since DNA helicase can unwind the double-stranded DNA without the need for heating, the HDA system provides a very useful tool to amplify DNA in vitro under isothermal conditions with a simplified reaction scheme. This review describes components and detailed aspects of current HDA systems using Escherichia coli UvrD helicase and T7 bacteriophage gp4 helicase with consideration of the processivity and efficiency of DNA amplification.
Jeong
Yong-Joo
YJ
Department of Bio and Nanochemistry, Kookmin University, Seoul, Republic of Korea.
Park
Kkothanahreum
K
Kim
Dong-Eun
DE
D017123:Bacteriophage T7 / Q000201:enzymology* / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008957:Models, Genetic; D021141:Nucleic Acid Amplification Techniques; D013696:Temperature
D029968:Escherichia coli Proteins; C506746:UvrD protein, E coli; D004265:DNA Helicases