Nucleotídeos para Mutagênese Randômica
Table 1: Nucleotide selection guide for random mutagenesis approaches.
Taq: Thermus aquaticus; transition: exchange of purine for purine (A ↔ G) or pyrimidine for pyrimidine (C ↔ T); transversion: exchange of purine for pyrimidine or vice versa (C/T ↔ A/G).
| Nucleotide | Method of incorporation | Primary type of mutation | Primarily induced mutation(s) | Reference |
|---|---|---|---|---|
| 8-Oxo-dGTP | PCR (Taq Pol) | Transversion | A:T → C:G and T:A → G:C | [1] |
| Combination of Error Prone-PCR (Mn2+) and PCR (Taq Pol) with 8-Oxo-dGTP | Transversion Transition | A:T → T:A and A:T → C:G A:T → G:C and G:C → A:T | [2] | |
| dPTP | PCR (Taq Pol) | Transition | A:T → G:C and G:C → A:T | [1] |
| 8-Oxo-dGTP & dPTP | PCR (Taq Pol) | Transversion Transition | Mixture of mutations induced by single nucleotides | [1,3] |
| 5Br-dUTP | PCR (Taq Pol) | Transversion Transition | A:T → G:C T:A → C:G | [4] |
| 2OH-dATP | PCR (Taq Pol) | Transversion Transition | A:T → C:G and G:C → T:A A:T → G:C | [5] |
| dITP | PCR (Taq Pol) | Transition | A:T → G:C and G:C → A:T | [6,7,8] |
Produtos
Selected References
[1] Zaccolo et al. (1996) An Approach to Random Mutagenesis of DNA Using Mixtures of Triphosphate Derivatives of Nucleoside Analogues. Journal of Molecular Biology 255:589.
[2] Kamiya et al. (2007) Induction of various mutations during PCRs with Manganese and 8-Hydroxy-dGTP. Biol. Pharm. Bull. 30 (4):842.
[3] Zaccolo et al. (1999) The effect of high-frequency random mutagenesis on in vitro protein evolution: a study on TEM-1 beta-lactamase. Journal of Molecular Biology 285 (2):775.
[4] Ma et al. (2008) The mutagenic properties of BrdUTP in a random mutagenesis process. Mol. Biol. Rep. 35:663.
[5] Kamiya et al. (2004) Induction of transition and transversion mutations during random mutagenesis PCR by the addition of 2-Hydroxy-dATP. Biol. Pharm. Bull. 27 (5):621.
[6] Kuipers (2012) In vitro mutagenesis by using mixtures of dNTP and dITP in PCR. In: Methods of Molecular Biology 57. Humana Press.
[7] Wang et al. (2012) A simple and reproducible method for direct evolution: Combination of random mutation with dITP and DNA fragmentation with Endonuclease V. In: Molecular Biotechnology. Humana Press.
[8] Spee et al. (1993) Efficient random mutagenesis method with adjustable mutation frequency using PCR and dITP. Nucleic Acids Res. 21 (3):777.