Difference between revisions of "Part:BBa K210009:Experience"

(→‎Applications of BBa_K210009)
(→‎Applications of BBa_K210009)
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'''iGEM Kyoto 2009'''
 
'''iGEM Kyoto 2009'''
  
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We mixed this part and liposome with fluorochrome and translated by RTS. When HIV-TAT::(LALAAAA)3 is translated, it sticks the lipid bilayer. As a result, the liposome which has HIV-TAT on their surface is completed. Adding it to HeLa cells. Later, we observed the fluorescence of liposome in HeLa cells by Confocal laser microscopy.
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To confirm the function of the constructed TAT-anchor peptide, we prepare the liposome containing NBD-PE
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with or without TAT-anchor which was synthesized or translated ''in vitro''.
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The size of the liposome without TAT was too small to distinguish individual particle, while that with TAT
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showed the large coalesced liposomes (Fig. NUMBER-(A)). The coalesced liposomes may be caused by the
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aggregation between TAT peptides or electric screening effect of TAT against liposome.
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Since the cells incubated with liposome did not emit any fluorescent signal, no liposome was uptaken by cells
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nonspecifically. In contrast, the TAT-liposome
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since if the liposome was uptaken by the
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cells, the fluorescent
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[[Image:Liposome-shape.png|700px|thumb|left|(A)Fluorescent images of liposome with or without TAT-anchor. (B)Microscopic images of the cells incubated with liposome. The green arrowheads indicated the positions of the fluorescent signals.]]
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[[Image:Liposome-transduction-slice.png|700px|thumb|left|Sliced cell treated with TAT-liposome. A merged image bright field and NBD-PE (left). The red dashed-lines indicate outline of cross-section of sliced cell. Red and green arrowheads are described in a text.]]
  
 
===User Reviews===
 
===User Reviews===

Revision as of 16:35, 21 October 2009

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Please enter how you used this part and how it worked out.

Applications of BBa_K210009

iGEM Kyoto 2009

To confirm the function of the constructed TAT-anchor peptide, we prepare the liposome containing NBD-PE with or without TAT-anchor which was synthesized or translated in vitro. The size of the liposome without TAT was too small to distinguish individual particle, while that with TAT showed the large coalesced liposomes (Fig. NUMBER-(A)). The coalesced liposomes may be caused by the aggregation between TAT peptides or electric screening effect of TAT against liposome. Since the cells incubated with liposome did not emit any fluorescent signal, no liposome was uptaken by cells nonspecifically. In contrast, the TAT-liposome since if the liposome was uptaken by the cells, the fluorescent

(A)Fluorescent images of liposome with or without TAT-anchor. (B)Microscopic images of the cells incubated with liposome. The green arrowheads indicated the positions of the fluorescent signals.


Sliced cell treated with TAT-liposome. A merged image bright field and NBD-PE (left). The red dashed-lines indicate outline of cross-section of sliced cell. Red and green arrowheads are described in a text.

User Reviews

UNIQb358863012b6eee5-partinfo-00000000-QINU UNIQb358863012b6eee5-partinfo-00000001-QINU