Polyelectrolyte surface interface for single-molecule fluorescence studies of DNA polymerase

被引：24

作者：

Kartalov, EP ^{[1
]}

Unger, MA ^{[1
]}

Quake, SR ^{[1
]}

机构：

[1] CALTECH, Dept Appl Phys, Pasadena, CA 91125 USA

来源：

BIOTECHNIQUES | 2003年 / 34卷 / 03期

关键词：

D O I：

10.2144/03343st02

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report the use of polyelectrolyte multilayers in a stable robust surface chemistry for specific anchoring of DNA to glass. The nonspecific binding of fluorescently tagged nucleotides is suppressed down to the single-molecule level, and DNA polymerase is active on the anchored DNA template. This surface-chemistry platform can be used for single-molecule studies of DNA and DNA polymerase and may be more broadly applicable for other situations in which it is important to have specific biomolecular surface chemistry with extremely low non-specific binding.

引用

页码：505 / +

页数：5

共 15 条

[1] Layer-by-layer assembled protein/polymer hybrid films: nanoconstruction via specific recognition
Cassier, T
Lowack, K
Decher, G
[J]. SUPRAMOLECULAR SCIENCE, 1998, 5 (3-4): : 309 - 315
[2] Single-molecule measurements calibrate green fluorescent protein surface densities on transparent beads for use with 'knock-in' animals and other expression systems
Chiu, CS
Kartalov, E
Unger, M
Quake, S
Lester, HA
[J]. JOURNAL OF NEUROSCIENCE METHODS, 2001, 105 (01) : 55 - 63
[3] Peptide hormone covalently bound to polyelectrolytes and embedded into multilayer architectures conserving full biological activity
Chluba, J
Voegel, JC
Decher, G
Erbacher, P
Schaaf, P
Ogier, J
[J]. BIOMACROMOLECULES, 2001, 2 (03) : 800 - 805
[4] Fuzzy nanoassemblies: Toward layered polymeric multicomposites
Decher, G
[J]. SCIENCE, 1997, 277 (5330) : 1232 - 1237
[5] PROOF OF MULTILAYER STRUCTURAL ORGANIZATION IN SELF-ASSEMBLED POLYCATION POLYANION MOLECULAR FILMS
DECHER, G
LVOV, Y
SCHMITT, J
[J]. THIN SOLID FILMS, 1994, 244 (1-2) : 772 - 777
[6] Initiation and re-initiation of DNA unwinding by the Escherichia coli Rep helicase
Ha, T
Rasnik, I
Cheng, W
Babcock, HP
Gauss, GH
Lohman, TM
Chu, S
[J]. NATURE, 2002, 419 (6907) : 638 - 641
[7] Single-molecule fluorescence spectroscopy of enzyme conformational dynamics and cleavage mechanism
Ha, TJ
Ting, AY
Liang, J
Caldwell, WB
Deniz, AA
Chemla, DS
Schultz, PG
Weiss, S
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (03) : 893 - 898
[8] Protein adsorption onto auto-assembled polyelectrolyte films
Ladam, G
Schaaf, P
Cuisinier, FJG
Decher, G
Voegel, JC
[J]. LANGMUIR, 2001, 17 (03) : 878 - 882
[9] Single-molecule enzymatic dynamics
Lu, HP
Xun, LY
Xie, XS
[J]. SCIENCE, 1998, 282 (5395) : 1877 - 1882
[10] ASSEMBLY OF THIN-FILMS BY MEANS OF SUCCESSIVE DEPOSITION OF ALTERNATE LAYERS OF DNA AND POLY(ALLYLAMINE)
LVOV, Y
DECHER, G
SUKHORUKOV, G
[J]. MACROMOLECULES, 1993, 26 (20) : 5396 - 5399

← 1 2 →