Other Publications (1)
Articles by Eliza L. Lewis in JoVE
アクリルアミドゲル電気泳動法で可視化した近赤外蛍光標識 DNA を用いた DNA ポリメラーゼ活性測定法 Eliza L. Lewis1, Aaron M. Leconte1 1Department of Chemistry, W.M. Keck Science Department of Claremont Mckenna, Pitzer, and Scripps College このプロトコルでは、電気泳動、ゲル画像を用いた近赤外蛍光に分類された DNA に変化の観察を通じて変更された DNA の DNA ポリメラーゼの合成の特性について説明します。アクリルアミドゲルはサイズに応じて異なる速度で移行短い核酸の分離の高分解能イメージングに使用されます。
Other articles by Eliza L. Lewis on PubMed
Design and Discovery of New Combinations of Mutant DNA Polymerases and Modified DNA Substrates Chembiochem : a European Journal of Chemical Biology. Apr, 2017 | Pubmed ID: 28160372 Chemical modifications can enhance the properties of DNA by imparting nuclease resistance and generating more-diverse physical structures. However, native DNA polymerases generally cannot synthesize significant lengths of DNA with modified nucleotide triphosphates. Previous efforts have identified a mutant of DNA polymerase I from Thermus aquaticus DNA (SFM19) as capable of synthesizing a range of short, 2'-modified DNAs; however, it is limited in the length of the products it can synthesize. Here, we rationally designed and characterized ten mutants of SFM19. From this, we identified enzymes with substantially improved activity for the synthesis of 2'F-, 2'OH-, 2'OMe-, and 3'OMe-modified DNA as well as for reverse transcription of 2'OMe DNA. We also evaluated mutant DNA polymerases previously only tested for synthesis for 2'OMe DNA and showed that they are capable of an expanded range of modified DNA synthesis. This work significantly expands the known combinations of modified DNA and Taq DNA polymerase mutants.