Difference between revisions of "Part:BBa K2986007"

 
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<partinfo>BBa_K2986007 short</partinfo>
 
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mRuby
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<h1>Usage and Biology:</h1>
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mRuby has the excitation and emission maxima at 558 nm and 605 nm, and a large Stokes shift of 47 nm, mRuby appears particularly useful for imaging applications.(Kredel, S., 2009). It is red fluorescent protein. This made the visualization of the target gene expression possible if we ligated the hGluc with target gene.
  
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[[File:T--SUSTech-enzyme.png|400px|thumb|center|Figure1.the plasmid used with hgluc]]
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===Usage and Biology===
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<h2>Design</h2>
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We wanted to design a gene expression system with mRuby as reporter, so that we can simplify the production detection process into fluorescent observation. We designed a plasmid ligate our target gene with mRuby using mammalian lentivirus expression vector. We use a 45bp linker to connect the hGluc with the CD11b transmembrane domain and use P2A (2A peptide allows an open reading frame (ORF) to translate a peptide chain into several independent peptide chains) connect with Interleukin 8 CDS, in our assumption this allow the stable expression of both our target gene and the hGluc in Hela cells.
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===Location of features===
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mRuby: 2379-3089
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hGluc: 3120-3710<br/>
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linker: 3711-3755<br/>
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CD11b transmembrane domain: 3756-3989<br/>
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P2A :3990-4055<br/>
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Interleukin 8 CDS :4056-4352<br/>
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<h1>Sequence and Features</h1>
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2986007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2986007 SequenceAndFeatures</partinfo>
  
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<h1>References</h1>
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Kredel, S., Oswald, F., Nienhaus, K., Deuschle, K., Röcker, C., Wolff, M., … Wiedenmann, J. (2009). mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures. PloS one, 4(2), e4391. doi:10.1371/journal.pone.0004391
  
 
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Revision as of 05:50, 19 October 2019


mRuby

Usage and Biology:

mRuby has the excitation and emission maxima at 558 nm and 605 nm, and a large Stokes shift of 47 nm, mRuby appears particularly useful for imaging applications.(Kredel, S., 2009). It is red fluorescent protein. This made the visualization of the target gene expression possible if we ligated the hGluc with target gene.

Figure1.the plasmid used with hgluc


Design

We wanted to design a gene expression system with mRuby as reporter, so that we can simplify the production detection process into fluorescent observation. We designed a plasmid ligate our target gene with mRuby using mammalian lentivirus expression vector. We use a 45bp linker to connect the hGluc with the CD11b transmembrane domain and use P2A (2A peptide allows an open reading frame (ORF) to translate a peptide chain into several independent peptide chains) connect with Interleukin 8 CDS, in our assumption this allow the stable expression of both our target gene and the hGluc in Hela cells.

Location of features

mRuby: 2379-3089 hGluc: 3120-3710
linker: 3711-3755
CD11b transmembrane domain: 3756-3989
P2A :3990-4055
Interleukin 8 CDS :4056-4352


Sequence and Features

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 16

References

Kredel, S., Oswald, F., Nienhaus, K., Deuschle, K., Röcker, C., Wolff, M., … Wiedenmann, J. (2009). mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures. PloS one, 4(2), e4391. doi:10.1371/journal.pone.0004391