Difference between revisions of "Part:BBa K2817013"
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===Usage and Biology===
 
===Usage and Biology===
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When testing the validity of the secretory tag YebF, we transformed the constructed YebF-GFP plasmid into DH5α and cultured overnight at 37 ℃. The supernatant was centrifuged at 3000 rpm for 5 min, and the supernatant was taken for fluorescence detection. The results are shown in the Figure 1. But we forgot to set up a positive control group. Due to time constraints, we have no time to repeat this experiment. But as a secretory tag used in both the literature and the iGEM team, we believe it can actually work.
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According to previous works, the secretory tag YebF has been demonstrated as the functioning part in E.coli，thus, we chose this tag for further experiment.
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When testing the validity of the secretory tag YebF, we transformed the constructed YebF-GFP plasmid into DH5α and cultured overnight at 37 ℃. The bacterial suspension was centrifuged at 3,000 r.p.m for 5 min and the supernatant was taken for fluorescence detection. The result is shown in the Figure 1. From our data, the YebF based secretory part worked efficiently.
  
 
https://static.igem.org/mediawiki/2018/9/92/T--NEU_China_A--results-6.png
 
https://static.igem.org/mediawiki/2018/9/92/T--NEU_China_A--results-6.png
  
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Figure 1. The fluorescence of overnight bacterial suspension after centrifugation at 3000 rpm for 5 min.
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Figure 6. The fluorescence density of overnight bacterial suspension.
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[1] Zhang G, Brokx S, Weiner JH (2006) Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli. Nat Biotechnol 24:100–104
  
 
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Latest revision as of 17:40, 17 October 2018

J23100-RBS-YebF-GFP

YebF is a secretion protein from E.coli. It’s used as secretion tag.We combine YebF with GFP to characterize its performance.

Usage and Biology

According to previous works, the secretory tag YebF has been demonstrated as the functioning part in E.coli，thus, we chose this tag for further experiment. When testing the validity of the secretory tag YebF, we transformed the constructed YebF-GFP plasmid into DH5α and cultured overnight at 37 ℃. The bacterial suspension was centrifuged at 3,000 r.p.m for 5 min and the supernatant was taken for fluorescence detection. The result is shown in the Figure 1. From our data, the YebF based secretory part worked efficiently.

T--NEU_China_A--results-6.png

Figure 6. The fluorescence density of overnight bacterial suspension.

[1] Zhang G, Brokx S, Weiner JH (2006) Extracellular accumulation of recombinant proteins fused to the carrier protein YebF in Escherichia coli. Nat Biotechnol 24:100–104

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1056