Difference between revisions of "Part:BBa K2986007"
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<h1>Usage and Biology:</h1> | <h1>Usage and Biology:</h1> | ||
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. | 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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===Location of features=== | ===Location of features=== | ||
| + | [[File:T--SUSTech-enzyme.png|400px|thumb|center|Figure1.the plasmid used with hgluc]] | ||
mRuby: 2379-3089 | mRuby: 2379-3089 | ||
hGluc: 3120-3710<br/> | hGluc: 3120-3710<br/> | ||
Latest revision as of 08:36, 20 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.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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