Difference between revisions of "Part:BBa K2666004"

 
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== VHH LaM-4 with signal peptid YncM with a L.jensenii specific strong promoter ==
  
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__NOTOC__
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=== I. Part BBa K2666000 : Function ===
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<partinfo>BBa_K2666004 short</partinfo>
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The Montpellier iGEM team 2018 designed a construction that produce and secrete LaM4, a nanobody against RFP and mcherry in Lactobacillus jensenii (more information). For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. The signal peptid is B. subtilis specific [2]. Figure 1 illustrates the detailed design.
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The Montpellier iGEM team 2018 designed a construction that produce and secrete LaM-4, a nanobody against RFP and mcherry in Lactobacillus jensenii (more information). For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. The signal peptid is B. subtilis specific [2]. Figure 1 illustrates the detailed design.
  
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Figure 1 : Construct design. The sequence that encode scFv LaM-4 with a SP YncM and produce by a strong promoter RpsU.
 
  
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LaM4 is used as a control [3]. It is an antibody against RFP & mcherry. This control allow us to check the efficiency of the promoter and the signal peptid.
 
  
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Proof of function
 
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WB
 
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ELISA Test
 
  
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Sequence and feature
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[[File:T--Montpellier--LaM-4-rpsU design mtp.PNG|550px]]
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seq on benchling
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Figure 1 : Construct design. That sequence encodes VHH LaM-4 with a SP YncM regulated by a strong promoter RpsU.
  
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LaM-4 is used as a control [3]. It is an antibody against RFP & mcherry. This control allow us to check the efficiency of the promoter and the signal peptid.
  
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=== II. Proof of function ===
  
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=== III. Design Considerations ===
  
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<!-- Add more about the biology of this part here
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We added spacers to all of our constructions to unable easier use of the sequence and separation of the different genes of the sequences. We used two Terminators to our sequences :BBa_B0014 & BBa_B0015 to ensure the stopping of the transcription.
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===Usage and Biology===
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== Reference: ==
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[1] Bao, Sujin, et al. "Distribution dynamics of recombinant Lactobacillus in the gastrointestinal tract of neonatal rats." PloS one 8.3 (2013): e60007.
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[2] Brockmeier, Ulf, et al. "Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria." Journal of molecular biology 362.3 (2006): 393-402.
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[3] Fridy, P. C., Li, Y., Keegan, S., Thompson, M. K., Nudelman, I., Scheid, J. F., ... & Rout, M. P. (2014). A robust pipeline for rapid prod
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2666004 parameters</partinfo>
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<partinfo>Part:BBa_K2666004 parameters</partinfo>
 
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Latest revision as of 12:53, 17 October 2018

VHH LaM-4 with signal peptid YncM with a L.jensenii specific strong promoter

I. Part BBa K2666000 : Function

The Montpellier iGEM team 2018 designed a construction that produce and secrete LaM-4, a nanobody against RFP and mcherry in Lactobacillus jensenii (more information). For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. The signal peptid is B. subtilis specific [2]. Figure 1 illustrates the detailed design.



T--Montpellier--LaM-4-rpsU design mtp.PNG

Figure 1 : Construct design. That sequence encodes VHH LaM-4 with a SP YncM regulated by a strong promoter RpsU.

LaM-4 is used as a control [3]. It is an antibody against RFP & mcherry. This control allow us to check the efficiency of the promoter and the signal peptid.

II. Proof of function

III. Design Considerations

We added spacers to all of our constructions to unable easier use of the sequence and separation of the different genes of the sequences. We used two Terminators to our sequences :BBa_B0014 & BBa_B0015 to ensure the stopping of the transcription.

Reference:

[1] Bao, Sujin, et al. "Distribution dynamics of recombinant Lactobacillus in the gastrointestinal tract of neonatal rats." PloS one 8.3 (2013): e60007.

[2] Brockmeier, Ulf, et al. "Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria." Journal of molecular biology 362.3 (2006): 393-402.

[3] Fridy, P. C., Li, Y., Keegan, S., Thompson, M. K., Nudelman, I., Scheid, J. F., ... & Rout, M. P. (2014). A robust pipeline for rapid prod

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 62
    Illegal BamHI site found at 194
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]