Separation efficiency of free-solution conjugated electrophoresis with drag-tags incorporating a synthetic amino acid
- Authors
- Seo, Kyung-Ho; Chu, Hun-Su; Yoo, Tae Hyeon; Lee, Sun-Gu; Won, Jong-In
- Issue Date
- Mar-2016
- Publisher
- WILEY-BLACKWELL
- Keywords
- Drag-tag; ELP; FSCE; GFP; Synthetic amino acid
- Citation
- ELECTROPHORESIS, v.37, no.5-6, pp.818 - 825
- Journal Title
- ELECTROPHORESIS
- Volume
- 37
- Number
- 5-6
- Start Page
- 818
- End Page
- 825
- URI
- https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/8056
- DOI
- 10.1002/elps.201500506
- ISSN
- 0173-0835
- Abstract
- DNA sequencing or separation by conventional capillary electrophoresis with a polymer matrix has some inherent drawbacks, such as the expense of polymer matrix and limitations in sequencing read length. As DNA fragments have a linear charge-to-friction ratio in free solution, DNA fragments cannot be separated by size. However, size-based separation of DNA is possible in free-solution conjugate electrophoresis (FSCE) if a "drag-tag" is attached to DNA fragments because the tag breaks the linear charge-to-friction scaling. Although several previous studies have demonstrated the feasibility of DNA separation by free-solution conjugated electrophoresis, generation of a monodisperse drag-tag and identification of a strong, site-specific conjugation method between a DNA fragment and a drag-tag are challenges that still remain. In this study, we demonstrate an efficient FSCE method by conjugating a biologically synthesized elastin-like polypeptide (ELP) and green fluorescent protein (GFP) to DNA fragments. In addition, to produce strong and site-specific conjugation, a methionine residue in drag-tags is replaced with homopropargylglycine (Hpg), which can be conjugated specifically to a DNA fragment with an azide site.
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Collections - College of Engineering > Chemical Engineering Major > 1. Journal Articles
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