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Volumn 91, Issue 6, 2015, Pages

Molecular behavior of DNA in a cell-sized compartment coated by lipids

Author keywords

[No Author keywords available]

Indexed keywords

CELLS; CONFORMATIONS; CYTOLOGY; FREE ENERGY; MEMBRANES; MOLECULES; PHOSPHOLIPIDS;

EID: 84936972905     PISSN: 15393755     EISSN: 15502376     Source Type: Journal    
DOI: 10.1103/PhysRevE.91.062717     Document Type: Article
Times cited : (16)

References (35)
  • 15
    • 84936965357 scopus 로고    scopus 로고
    • We prepared 2.0 ng/mL T4 DNA (Nippon Gene) with YOYO-1 iodide (Invitrogen) (0.1 dye/bp), 10 mM Tris-HCl (pH 7.4), and various concentrations of SPD (SERVA Electrophoresis GmbH). DNA and YOYO-1 were first mixed and incubated to reach equilibrium, and then Tris-HCl and SPD were added to obtain the DNA solution with final concentrations. The aqueous solution was dispersed in olive oil (Wako Pure Chemicals) containing dioleoyl L-(Equation presented) phosphatidylcholine (DOPC) (Avanti Polar Lipids) to obtain cell-sized water-in-oil droplets (the detailed procedure have been reported previously [16]). T4 DNA in the droplet was observed using a laser scanning microscope (FV1000-D, Olympus). Single molecules of T4 DNA were also observed using a fluorescent microscope (IX71, Olympus) equipped with a confocal scanner (CSU-10, Yokogawa)
    • We prepared 2.0 ng/mL T4 DNA (Nippon Gene) with YOYO-1 iodide (Invitrogen) (0.1 dye/bp), 10 mM Tris-HCl (pH 7.4), and various concentrations of SPD (SERVA Electrophoresis GmbH). DNA and YOYO-1 were first mixed and incubated to reach equilibrium, and then Tris-HCl and SPD were added to obtain the DNA solution with final concentrations. The aqueous solution was dispersed in olive oil (Wako Pure Chemicals) containing dioleoyl L-(Equation presented) phosphatidylcholine (DOPC) (Avanti Polar Lipids) to obtain cell-sized water-in-oil droplets (the detailed procedure have been reported previously [16]). T4 DNA in the droplet was observed using a laser scanning microscope (FV1000-D, Olympus). Single molecules of T4 DNA were also observed using a fluorescent microscope (IX71, Olympus) equipped with a confocal scanner (CSU-10, Yokogawa).
  • 17
    • 84936965358 scopus 로고    scopus 로고
    • Notably, this results cannot be attributed to the time-scale required to achieve an equilibrium configuration, since DNA in small droplets can contact the membrane surface more rapidly by diffusion
    • Notably, this results cannot be attributed to the time-scale required to achieve an equilibrium configuration, since DNA in small droplets can contact the membrane surface more rapidly by diffusion.
  • 18
    • 84936965359 scopus 로고    scopus 로고
    • To construct the phase diagram, we counted the number of droplets with adsorbed DNA (0, 0.2, and 0.5 mM SPD) and unfolded DNA (1.0 and 2.5 mM SPD) to obtain the probabilities as a function of droplet size for each SPD concentration [see also Fig. 4(c) and 4(d)]. The total number of droplets is (Equation presented) (0.2 mM SPD), (Equation presented) (0.5 mM SPD), (Equation presented) (1.0 mM SPD), and (Equation presented) (2.5 mM SPD) from two or three repeats of the counting
    • To construct the phase diagram, we counted the number of droplets with adsorbed DNA (0, 0.2, and 0.5 mM SPD) and unfolded DNA (1.0 and 2.5 mM SPD) to obtain the probabilities as a function of droplet size for each SPD concentration [see also Fig. 4(c) and 4(d)]. The total number of droplets is (Equation presented) (0.2 mM SPD), (Equation presented) (0.5 mM SPD), (Equation presented) (1.0 mM SPD), and (Equation presented) (2.5 mM SPD) from two or three repeats of the counting.
  • 19
    • 84936965360 scopus 로고    scopus 로고
    • Parameter: (Equation presented) m, (Equation presented), (Equation presented), (Equation presented), (Equation presented), (Equation presented), and (Equation presented)
    • Parameter: (Equation presented) m, (Equation presented), (Equation presented), (Equation presented), (Equation presented), (Equation presented), and (Equation presented).
  • 20
    • 0017895903 scopus 로고
    • G. S. Manning, Q. Rev. Biophys. 11, 179 (1978). QURBAW 0033-5835 10.1017/S0033583500002031
    • (1978) Q. Rev. Biophys. , vol.11 , pp. 179
    • Manning, G.S.1
  • 22
    • 84936965361 scopus 로고    scopus 로고
    • We measured the (Equation presented) potential of DOPC liposomes with 10 mM Tris-HCl (pH 7.4) using a Zetasizer Nano ZS (Malvern Instruments, Worcestershire, United Kingdom) at 25 (Equation presented)
    • We measured the (Equation presented) potential of DOPC liposomes with 10 mM Tris-HCl (pH 7.4) using a Zetasizer Nano ZS (Malvern Instruments, Worcestershire, United Kingdom) at 25 (Equation presented).
  • 31
    • 74849118341 scopus 로고    scopus 로고
    • D. Lingwood and K. Simons, Science 327, 46 (2010). SCIEAS 0036-8075 10.1126/science.1174621
    • (2010) Science , vol.327 , pp. 46
    • Lingwood, D.1    Simons, K.2
  • 35
    • 84936965363 scopus 로고    scopus 로고
    • Dipalmitoyl L-(Equation presented) phosphatidylcholine (DPPC), cholesterol (Chol) (Avanti Polar Lipids), and N-(rhodamine red-X)-1, 2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethyl-am-monium salt (rho-DHPE) (Invitrogen) were used. We tested several lipid mixtures (DOPC/DPPC/cholesterol=0/50/50, 50/0/50, 20/60/20) to produce order-disorder lateral phase separation, as well as another fluorescent dye Texas-red DHPE. There was no apparent difference in the phase partitioning of DNA molecules among these membrane materials
    • Dipalmitoyl L-(Equation presented) phosphatidylcholine (DPPC), cholesterol (Chol) (Avanti Polar Lipids), and N-(rhodamine red-X)-1, 2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethyl-am-monium salt (rho-DHPE) (Invitrogen) were used. We tested several lipid mixtures (DOPC/DPPC/cholesterol=0/50/50, 50/0/50, 20/60/20) to produce order-disorder lateral phase separation, as well as another fluorescent dye Texas-red DHPE. There was no apparent difference in the phase partitioning of DNA molecules among these membrane materials.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.