Difference between revisions of "Part:BBa K2984030"

 
 
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<partinfo>BBa_K2984030 short</partinfo>
 
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L1 construc containg Arylsulfatase secretion Signal, mVenus fluorescent marker, SP20 glycomodule. These parts are flanked by the Psad Promoter and the Rbcs2 Terminator. The construct can be used for secretion analysis in C.reinhardtii. The glycomodule is supposed to ramp up secretion and post translational glycosilations.  
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This vector is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. L1 construct containg arylsulfatase secretion Signal <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984000">ARS</a>, <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984020">mVenus</a> fluorescent marker, <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984024">SP20</a> glycomodule. These parts are flanked by the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984008">PsaD promoter</a> and the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984021">Rbcs2 terminator</a>. The construct can be used for secretion analysis in <i>C.reinhardtii</i>. The glycomodule is supposed to ramp up secretion and post translational glycosilations.  
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<partinfo>BBa_K2984030 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2984030 SequenceAndFeatures</partinfo>
  
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==Characterization==
  
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We were able to demonstrate the presence of YFP mVenus in <i>C. reinhardtii</i> transformed with this construct. <i>C. reinhardtii</i> shows vesicles near to the membrane area containing high concentrations of YFP mVenus, which indicates the secretion mechanism driven by the secretion signal ARS and the glycomodule SP<sub>20</sub>.
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===Functional Parameters===
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<partinfo>BBa_K2984030 parameters</partinfo>
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<img class="is-revealing" src="https://2019.igem.org/wiki/images/b/b2/T--Humboldt_Berlin--Microscopy-YFP.png" alt="fluorescence difference spectrum" width="600" />
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<figcaption>Fig. 1 - Presence of YFP mVenus in <i>C. reinhardtii</i>. For construct BBa K2984030 it can be seen that a vesicle with a high concentration of YFP mVenus was formed near the membrane. Taken with a confocal microscope.</figcaption>
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Latest revision as of 21:47, 21 October 2019


L1c-Psad-ARS-YFP-SP20-Rbcs2

This vector is a part of the Chlamy-HUB-Collection. L1 construct containg arylsulfatase secretion Signal ARS, mVenus fluorescent marker, SP20 glycomodule. These parts are flanked by the PsaD promoter and the Rbcs2 terminator. The construct can be used for secretion analysis in C.reinhardtii. The glycomodule is supposed to ramp up secretion and post translational glycosilations.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1110
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1110
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1110
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1110
    Illegal NgoMIV site found at 1587
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

We were able to demonstrate the presence of YFP mVenus in C. reinhardtii transformed with this construct. C. reinhardtii shows vesicles near to the membrane area containing high concentrations of YFP mVenus, which indicates the secretion mechanism driven by the secretion signal ARS and the glycomodule SP20.

fluorescence difference spectrum

Fig. 1 - Presence of YFP mVenus in C. reinhardtii. For construct BBa K2984030 it can be seen that a vesicle with a high concentration of YFP mVenus was formed near the membrane. Taken with a confocal microscope.