Team:NYMU-Taipei/results/immunological-solution1

From 2011.igem.org

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'''<font size=4><font color=blue>Related Parts</font>'''</font>
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<font size=3>We have created several PARTs on the basis of Biobrick. The expression will be shown on the partregistry webpage.
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{| border=1
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|rowspan=1 |  [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624033 '''<u>BBa_K624033</u>''']
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|minC cell division inhibitor (revised)
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|-
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|rowspan=1 |  [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624034 '''<u>BBa_K624034</u>''']
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|revised minC primer (forward)
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|-
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|rowspan=1 |  [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624035 '''<u>BBa_K624035</u>''']
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|revised minC primer (reverse)
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624036 '''<u> BBa_K624036</u>''']
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|minC+mcherry
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624037 '''<u>BBa_K624037</u>''']
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| rbs+minC+mcherry
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624040 '''<u>BBa_K624040</u>''']
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|pT7-tetO+RBS+minC
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624045 '''<u>BBa_K624045</u>''']
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| Pmsp1(tetO) + RBS(Pmsp3) + minC + RBS(Pmsp3) + mcherry
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624056 '''<u>BBa_K624056</u>''']
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| pYMB essentials + RBS(Pmsp3) + minC
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624057 '''<u>BBa_K624057</u>''']
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| pYMB essentials + RBS(Pmsp3) + LLO
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|-
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| rowspan=1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K624058 '''<u>BBa_K624058</u>''']
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| pYMB essentials + RBS(Pmsp3) + Inv
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|-
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|}
===<font size=4><font color=blue>'''Nested-PCR Primers For Various Strains'''</font></font>===
===<font size=4><font color=blue>'''Nested-PCR Primers For Various Strains'''</font></font>===

Revision as of 03:36, 6 October 2011

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Contents

Construction & Parts

Our minC Construct and Inv & LLO Construct on AMB-1

BBa_K624053


AMB-1 minC NYMU.jpg
AMB-1 Inv LLO.jpg
Fig. Design of minC Construct on AMB-1
Backbone Plasmid: pYMB
Fig. Design of Inv & LLO Construct on AMB-1
Backbone Plasmid: pRKm415

Related Parts We have created several PARTs on the basis of Biobrick. The expression will be shown on the partregistry webpage.

BBa_K624033 minC cell division inhibitor (revised)
BBa_K624034 revised minC primer (forward)
BBa_K624035 revised minC primer (reverse)
BBa_K624036 minC+mcherry
BBa_K624037 rbs+minC+mcherry
BBa_K624040 pT7-tetO+RBS+minC
BBa_K624045 Pmsp1(tetO) + RBS(Pmsp3) + minC + RBS(Pmsp3) + mcherry
BBa_K624056 pYMB essentials + RBS(Pmsp3) + minC
BBa_K624057 pYMB essentials + RBS(Pmsp3) + LLO
BBa_K624058 pYMB essentials + RBS(Pmsp3) + Inv

Nested-PCR Primers For Various Strains

nested-Inv

BBa_K624063 nested_Inv F (inter-strain nested primer)
BBa_K624064 nested_Inv R (inter-strain nested primer)

nested-LLO

BBa_K624065 nested_LLO F (inter-strain nested primer)
BBa_K624066 nested_LLO R (inter-strain nested primer)

Symbiosis with Human Glial Cell

Magnetotactic bacteria were introduced into guanulocytes and monocytes by phagocytosis.(Tadashi Matsunaga et all. 1986) Monocyte.jpg

In order to simulate the circumstances in brain, we put AMB-1 separately into mixed glial cells in vitro. Below are the photos we captured under microscope.

check out BBa_K624032 for detailed characterization

File:AMB-1 glial1.tif File:AMB-1 glial2.tif
File:Glial1.tif File:Glial2.tif

Measurements of Construct tetR+rbs+gfp under Fluorescence Microscope

30.JPG 31.JPG 32.JPG


These are the measurements of our construct tetR+rbs+GFP(BBa_K624001) under fluorescence microscope. The left graph is from the control group of E. coli strain DH5-alpha. The middle is tetR+rbs+gfp in plasmid pSB1C3 in E. coli, while the right is from a plate without any bacilli. All graphs are measured in fixed exposure time. Construct of tetR+rbs+gfp which emits light is thought to be a leak phenomenon of plasmid, while RE digest has proved that the sequence length is correct, and further sequencing work is under operation.


gfp measurements under different excitation wavelengths


33.JPG


The expression of gfp is measured precisely under excitation wavelength from 475nm to 501nm, given the absorption wavelength 511nm. In our measurement, wavelength over 493nm yields high background, resulting in values of control group surging far beyond normal condition. Thus, 493nm is suggested as the best excitation wavelength, as it presents the highest emission light and the lowest of the control group.