Publication

Investigation of various fluorescent protein-DNA binding peptides for effectively visualizing large DNA molecules

조회수 : 167 등록일 : 2016.07.14 00:00

저자 : Lee, S (Lee, Seonghyun), Lee, HS (Lee, Hyun Soo), Jo, K (Jo, Kyubong)
출처 : RSC ADVANCES
출판일 : 2016.07.01
 
 
Investigation of various fluorescent protein-DNA binding peptides for effectively visualizing large DNA molecules
 
 
Lee, S (Lee, Seonghyun)[ 1,2 ] ; Wang, C (Wang, Cong)[ 3 ] ; Song, J (Song, Junghyun)[ 1,2 ] ; Kim, DG (Kim, Do-geun)[ 1,2 ] ; Oh, Y (Oh, Yeeun)[ 1,2 ] ; Ko, W (Ko, Wooseok)[ 1,2 ] ; Lee, J (Lee, Jinyong)[ 1,2 ] ; Park, J (Park, Jungyul)[ 3 ] ; Lee, HS (Lee, Hyun Soo)[ 1,2 ] ; Jo, K (Jo, Kyubong)[ 1,2 ]
 
 
[ 1 ] Sogang Univ, Dept Chem, 1 Shinsudong, Seoul 121742, South Korea
[ 2 ] Sogang Univ, Program Integrated Biotech, 1 Shinsudong, Seoul 121742, South Korea
[ 3 ] Sogang Univ, Dept Mech Engn, 1 Shinsudong, Seoul 121742, South Korea
 
 
 
Large DNA molecules were visualized with novel fluorescent protein-DNA binding peptides (FP-DBPs). We constructed FP-DBPs by linking fluorescent protein to the N- or C-terminal of one or two peptides. We designed these DNA binding peptides from various DNA binding motifs such as oligo-lysine (K) and LysTrp (KW) repeats, TPKRPRGRPKK from high mobility group (HMG) chromosomal protein, and Ser-ProArg- Lys (SPRK) from histone protein. We demonstrated the use of FP-DBP to stain large DNA molecules, and then analysed the fluorescence brightness and their binding affinity to double stranded DNA. This investigation provided HMG-tagged FP-DBP as the best DNA staining reagent in terms of fluorescence intensity, signal-to-noise ratio, and DNA binding affinity (K-d = 586 nM). Furthermore, we measured elongation of FP-DBP-stained DNA molecules tethered on the surface in order to evaluate FP-DBP-induced structural deformation.
 
 
 
 
 
 
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