Single-molecule DNA visualization using AT-specific red and non-specific green DNA-binding fluorescent proteins
- Authors
- Park, Jihyun; Lee, Seonghyun; Won, Nabin; Shin, Eunji; Kim, Soo-Hyun; Chun, Min-Young; Gu, Jungyeun; Jung, Gun-Young; Lim, Kwang-Il; Jo, Kyubong
- Issue Date
- Feb-2019
- Publisher
- ROYAL SOC CHEMISTRY
- Citation
- ANALYST, v.144, no.3, pp 921 - 927
- Pages
- 7
- Journal Title
- ANALYST
- Volume
- 144
- Number
- 3
- Start Page
- 921
- End Page
- 927
- URI
- https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/3835
- DOI
- 10.1039/c8an01426d
- ISSN
- 0003-2654
1364-5528
- Abstract
- The recent advances in the single cell genome analysis are generating a considerable amount of novel insights into complex biological systems. However, there are still technical challenges because each cell has a single copy of DNA to be amplified in most single cell genome analytical methods. In this paper, we present a novel approach to directly visualize a genomic map on a large DNA molecule instantly stained with red and green DNA-binding fluorescent proteins without DNA amplification. For this visualization, we constructed a few types of fluorescent protein-fused DNA-binding proteins: H-NS (histone-like nucleoid-structuring protein), DNA-binding domain of BRCA1 (breast cancer 1), high mobility group-1 (HMG), and lysine tryptophan (KW) repeat motif. Because H-NS and HMG preferentially bind A/T-rich regions, we combined A/T specific binder (H-NS-mCherry and HMG-mCherry as red color) and a non-specific complementary DNA binder (BRCA1-eGFP and 2(KW)(2)-eGFP repeat as green color) to produce a sequence-specific two-color DNA physical map for efficient optical identification of single DNA molecules.
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