Difference between revisions of "Part:BBa K2729006"
 
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EGFP contains chromophore mutations that make the protein 35 times brighter than wild-type GFP, and is codon-optimized for higher expression in mammalian cells.
 
EGFP contains chromophore mutations that make the protein 35 times brighter than wild-type GFP, and is codon-optimized for higher expression in mammalian cells.
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FMDV 2A is used widely for this purpose since multiple proteins can be coexpressed by linking open reading frames (ORFs) to form a single cistron.
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FMDV 2A is "self-cleaving" peptides, and upstream coding region can be a single cistron with the assistance.
  
 
===Characterization===
 
===Characterization===
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EGFP is derived from <i>the jellyfish Aequorea victoria</i> and optimized for <i>Homo Sapiens</i> using IDT Codon Optimization Tool. Our team checked the sequence length of synthesized oligo DNA and characterized it.
 
EGFP is derived from <i>the jellyfish Aequorea victoria</i> and optimized for <i>Homo Sapiens</i> using IDT Codon Optimization Tool. Our team checked the sequence length of synthesized oligo DNA and characterized it.
  
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[[File:T--Tokyo_Tech--test-electrop.gif|thumb|left|300px| '''Figure 1:''' '''Result of the qualitative experiment''' - description comes here. ]]
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[[File:Tokyotech electrophoresis.png|thumb|left|300px| '''Figure 1:''' '''Result of the Electrophoresis (Marker: lambda HindIII)''' ]]
  
 
<br>
 
<br>
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As shown in Figure 1, the band comes between 900 bp to 1,000 bp.  
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As shown in Figure 1, the band comes between 900 bp to 1,000 bp. The actual length without prefix and suffix is 950 bp.
 
<br style="clear: both" />
 
<br style="clear: both" />
  
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Our team successfully validated that the sequence was synthesized, and obtained enough amount of it for transfection.
 
Our team successfully validated that the sequence was synthesized, and obtained enough amount of it for transfection.
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The function of FMDV2a will be tested by testing the infection of pseudo-virus into Vero cells. If the 2a peptide works well, the fluorescence intensity can be observed robustly.
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===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K2729006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2729006 SequenceAndFeatures</partinfo>
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===Reference===
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- [https://www.chromotek.com/products/egfp/ EGFP - ChromoTek]

Latest revision as of 03:16, 18 October 2018


DENV1 ancC-prM-E (Dengue Virus Serotype I)

Usage and Biology

EGFP contains chromophore mutations that make the protein 35 times brighter than wild-type GFP, and is codon-optimized for higher expression in mammalian cells. FMDV 2A is "self-cleaving" peptides, and upstream coding region can be a single cistron with the assistance.

Characterization

EGFP is derived from the jellyfish Aequorea victoria and optimized for Homo Sapiens using IDT Codon Optimization Tool. Our team checked the sequence length of synthesized oligo DNA and characterized it.

Figure 1: Result of the Electrophoresis (Marker: lambda HindIII)


As shown in Figure 1, the band comes between 900 bp to 1,000 bp. The actual length without prefix and suffix is 950 bp.

Analysis

Our team successfully validated that the sequence was synthesized, and obtained enough amount of it for transfection.

The function of FMDV2a will be tested by testing the infection of pseudo-virus into Vero cells. If the 2a peptide works well, the fluorescence intensity can be observed robustly.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

- EGFP - ChromoTek