Difference between revisions of "Part:BBa K2666004"

 
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== scFv LaM4 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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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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WB
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ELISA Test
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Sequence and feature
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seq on benchling
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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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=== II. Proof of function ===
  
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=== III. Design Considerations ===
  
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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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== Reference: ==
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===Usage and Biology===
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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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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2666004 SequenceAndFeatures</partinfo>
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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===
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<partinfo>BBa_K2666004 parameters</partinfo>
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Revision as of 08:18, 10 October 2018

scFv LaM4 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.


Figure 1 : Construct design. The sequence that encode scFv LaM-4 with a SP YncM and produce by a strong promoter RpsU.

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.

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