Difference between revisions of "Team:Tokyo Tech"

(Undo revision 174708 by Bank-Tharachai (talk))
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      <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px" align="center">Dengue Virus Detection System with multiple colors</h1>
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      <h4 style="text-align: center">Virus detection system with multi-colors</h4>
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      <p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px">
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      For 2018 project, we try to develop mosaic-envelope virus that can detect the presence of antibodies in serum that correspond to specific type of virus. This mosaic-envelope virus can detect all the serotypes of dengue viruses and can contribute to versatile dengue virus detection. The experimental procedures are categorized as the following three steps.
 +
      </p>
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      <p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px">
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        STEP1: We construct integrated plasmid that codes essential viral proteins (both structural and non-structural) and green fluorescence protein. (Note: If the size is too large, they will be separated.) The envelope (E) genes are derived from 2 or more dengue serotypes so that envelope proteins can be mixed to detect multiple types of antibodies.
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      </p>
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      <p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px">
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      STEP2: We transduce the plasmid into suitable eukaryotic cells and culture the cells which are successfully transduced. (Drug selection and a series of passage may be needed.) Most of the transduced cells are expected to produce mosaic-envelope virus.
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      </p>
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      <p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px">
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      <p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px">
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        STEP3: We culture the target mammalian cells that are for checking the infection by mosaic-envelope virus. After mixing the cells and the virus, we measure the fluorescence intensity to check whether the mosaic-envelope virus infected and GFPs are produced or not. As control experiment, we add antibodies that correspond to one of the dengue serotypes to check whether the function of the virus is inhibited or not.
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      <figcaption style="font-size: 16px">Fig. 1 Basic Flow of dengue virus detection system</figcaption>
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Revision as of 06:56, 28 September 2018

Coli Sapiens

Project Description

Dengue Virus Detection System with multiple colors

Mechanism

For 2018 project, we try to develop mosaic-envelope virus that can detect the presence of antibodies in serum that correspond to specific type of virus. This mosaic-envelope virus can detect all the serotypes of dengue viruses and can contribute to versatile dengue virus detection. The experimental procedures are categorized as the following three steps.

STEP1: We construct integrated plasmid that codes essential viral proteins (both structural and non-structural) and green fluorescence protein. (Note: If the size is too large, they will be separated.) The envelope (E) genes are derived from 2 or more dengue serotypes so that envelope proteins can be mixed to detect multiple types of antibodies.

STEP2: We transduce the plasmid into suitable eukaryotic cells and culture the cells which are successfully transduced. (Drug selection and a series of passage may be needed.) Most of the transduced cells are expected to produce mosaic-envelope virus.

STEP3: We culture the target mammalian cells that are for checking the infection by mosaic-envelope virus. After mixing the cells and the virus, we measure the fluorescence intensity to check whether the mosaic-envelope virus infected and GFPs are produced or not. As control experiment, we add antibodies that correspond to one of the dengue serotypes to check whether the function of the virus is inhibited or not.

Fig. 1 Basic Flow of dengue virus detection system



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 iGEM Tokyo Tech

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Dengue Virus Detection System with multiple colors


Mechanism


Virus detection system with multi-colors

For 2018 project, we try to develop mosaic-envelope virus that can detect the presence of antibodies in serum that correspond to specific type of virus. This mosaic-envelope virus can detect all the serotypes of dengue viruses and can contribute to versatile dengue virus detection. The experimental procedures are categorized as the following three steps.

STEP1: We construct integrated plasmid that codes essential viral proteins (both structural and non-structural) and green fluorescence protein. (Note: If the size is too large, they will be separated.) The envelope (E) genes are derived from 2 or more dengue serotypes so that envelope proteins can be mixed to detect multiple types of antibodies.

STEP2: We transduce the plasmid into suitable eukaryotic cells and culture the cells which are successfully transduced. (Drug selection and a series of passage may be needed.) Most of the transduced cells are expected to produce mosaic-envelope virus.

<p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px"> STEP3: We culture the target mammalian cells that are for checking the infection by mosaic-envelope virus. After mixing the cells and the virus, we measure the fluorescence intensity to check whether the mosaic-envelope virus infected and GFPs are produced or not. As control experiment, we add antibodies that correspond to one of the dengue serotypes to check whether the function of the virus is inhibited or not.

     </p>
     <figure>
     <img src="T--Tokyo_Tech--virus_detec_flow_0629.png" style="max-width:80%">
     <figcaption style="font-size: 16px">Fig. 1 Basic Flow of dengue virus detection system</figcaption>
     </figure>

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