Biomod/2011/Tianjin:Results
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Project
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Contents
1
Results
1.1
CFS with microfluidics
1.2
CFS with SWNTs
1.2.1
AFM
1.2.2
SEM
1.2.3
TEM
1.2.4
Fluorescence micrograph
Results
CFS with microfluidics
Fig 1. Cell-free protein expression in W/O/W. Fluorescence micrograph of a W/O/W during the expression of (a) RFP as a soluble protein in the protocell cytosol, and (b) MreB-RFP, which has portioned at the hydrophobic interface. Figure (c) shows a confocal micrograph (3um thick plane) of a W/O/W, 14 h after generation. The RFP-tagged MreB polymerized, forming aggregates localized to the oil-water interface. The image was created by forming the W/O/W droplets with 100 nM BODIPY dispersed in the oil phase and MreB-RFP in the segmented phase. Fluorescence was detected in separate channels.
CFS with SWNTs
AFM
File:Image:Biomod tianjin results 11.png
Fig 2.AFM analysis of MreB-RFP cell-free aggregation with SWNTs, indicating that SWNTs serve as scaffolds in the spiral winding of fusion proteins in vitro. a). AFM image of fusion protein MreB-RFP without SWNT scaffold, protein aggregated in disorder and only discrete spherical particles were formed. b). AFM image of fusion protein MreB-RFP at the presence of SWNT, protein aggregated and wound spirally onto the surface of nanotube.
SEM
Fig 3. SEM analysis of MreB-RFP cell-free aggregation with SWNTs. a). SEM image of SWNT, the sidewall of nanotube is smooth without any absorption or aggregation. b). SEM image of MreB-RFP without scaffold, protein aggregated in disorder and only discrete spherical particles were formed. c). SEM image of fusion protein at the presence of SWNT, protein aggregated and wound spirally onto the surface of nanotube.
TEM
Fig 4. TEM analysis of MreB-RFP cell-free aggregation with SWNTs. a). TEM image of SWNT, the sidewall of nanotube is smooth without any absorption or aggregation. b). TEM image of MreB-RFP without scaffold, protein aggregated in disorder and only discrete spherical particles were formed. c). TEM image of fusion protein without SWNT scaffold, protein aggregated and wound spirally onto the surface of SWNT.
Fluorescence micrograph
Fig 5. Fluorescence micrograph of fusion protein MreB combined with SWNT. Red fluorescence can be observed, indicating the activity of fusion protein were reserved.
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