Difference between revisions of "Team:NTU-LIHPAO-Taiwan/Project/Parts"

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<img src="https://static.igem.org/mediawiki/2015/3/34/NTU-Team-nisI.jpg" width="480px"/>
 
<img src="https://static.igem.org/mediawiki/2015/3/34/NTU-Team-nisI.jpg" width="480px"/>
<div class="Intro_PictureText"><div class="Text_Picture">Fig.2-1-1 Nisin Selection</div></div>
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<div class="Intro_PictureText"><div class="Text_Picture">Fig.2-1 Nisin Selection</div></div>
 
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<img src="https://static.igem.org/mediawiki/2015/6/6a/NTU-Team-tat-pyy-his.jpg" width="480px"/>
<div class="Intro_PictureText"><div class="Text_Picture">Fig.2-3-1 Suicide Kill Part</div></div>
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<img src="https://static.igem.org/mediawiki/2015/4/4d/NTU-Team-plac-gfp.jpg" width="480px"/>
<div class="Intro_PictureText"><div class="Text_Picture">Fig.3-1-1 Cell Penetrating Peptide + PYY</div></div>
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<div class="Intro_PictureText"><div class="Text_Picture">Fig.3-1 Cell Penetrating Peptide + PYY</div></div>
 
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<img src="https://static.igem.org/mediawiki/2015/1/12/NTU-Team-his-pyy.jpg" width="480px"/>
<div class="Intro_PictureText"><div class="Text_Picture">Fig.3-2-1 Suicide Sense TestY</div></div>
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<img src="https://static.igem.org/mediawiki/2015/3/39/NTU-Team-sp-pyy-his.jpg" width="480px"/>
<div class="Intro_PictureText"><div class="Text_Picture">Fig.3-3-1 PYY</div></div>
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<img src="https://static.igem.org/mediawiki/2015/e/ea/NTU-Team-y2r.jpg" width="480px"/>
<div class="Intro_PictureText"><div class="Text_Picture">Fig.3-4-1 Signal peptide + PYY</div></div>
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Revision as of 06:22, 7 September 2015

NTU-LIHPAO-Taiwan

Lactobacillus casei
Prototype
Prototype
The iGEM Team NTU-LIHPAO-Taiwan 2015 had built a new biological system for the iGEM community. The original design of the system making up the (Product) contains the three elements listed below, in a probiotic – Lactobacillus casei ATCC 393.
Main Part
Part: BBa_K1841001
nisI
Research from Torsten et al. revealed that the highest level of acquired nisin tolerance was achieved after coordinated expression of all four nisin immunity genes, containing nisI, nisF, nisE, and nisG.[1] Functional analyses provided evidence that NisI acts as a nisin-sequestering protein and that NisFEG acts as a nisin exporter that expels nisin molecules from the cytoplasmic membrane into the environment.
Fig.2-1 Nisin Selection
Part: BBa_K1841004
plac promoter
Lactose operon of Lactobacillus casei In Lactobacillus casei ATCC393 [pLZ15–], the lactose genes are grouped in a cluster transcribed as single operon. The cluster lacTEGF encodes an antiterminator protein (LacT), lactose-specific elements (LacE and LacF) of the phosphotransferase system (PTS) and a phospho-β-galactosidase (LacG). The promoter region contains a cre element (catabolite responsive element) overlapping the –35 region, which is followed by a highly conserved sequence, the ribonucleic antiterminator (RAT) sequence, and a terminator structure. The antiterminator activity of LacT is also negatively controlled by glucose, possibly by PTS-mediated phosphorylation as explained below.[2]
Fig.2-2-1 Suicide Sense Part
Fig.2-2-2 Suicide Kill Part
Part: BBa_K1841007
TAT-PYY
TAT can bring the big molecule substance like DNA, protein, peptide to penetrate the cell. Usually, named as protein transduction domain or membrane transduction domain, it is less than 40 amino acid.[3] PYY belongs to the gastrointestinal hormones. Once the intestine detects the nutrition, the epidermal cell, L cell of ileum and colon will secrete the PYY. While the blood pass the hypothalamus, PYY will bind to the neuropeptide Y receptor in ventromedial nuclei causing the sense of satiation.[4]
Fig.2-3 Suicide Kill Part
In this part, we use the constitutive promoter BBa_J23100 to produce signal peptide-PYY-histidine fusion protein. With the His-tag, we can easily purify the signal peptide-PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY that can secret out of the L. casei.
Test Part
RFP
Fig.3-1 Cell Penetrating Peptide + PYY
In this part, we use the RFP protein to testify the function of the lactose operon.
His-PYY
Fig.3-2 Suicide Sense TestY
In this part, we use the constitutive promoter BBa_J23100 to produce histidine-PYY fusion protein. With the His-tag, we can easily purify the PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY under the constitutive promoter in the L. casei.
Signal peptide-PYY-His
Fig.3-3 PYY
In this part, we use the constitutive promoter BBa_J23100 to produce signal peptide-PYY-histidine fusion protein. With the His-tag, we can easily purify the signal peptide-PYY from other non-His-tag protein. This part is constructed to test the amount of the PYY that can secret out of the L. casei.
Y2R
Fig.3-4 Signal peptide + PYY
In this part, we use the constitutive promoter BBa_J23100 to produce Y2R. This part is constructed to verify the function of PYY.
Reference
Reference
[1] Torsten, S., Stefan, H., Irina, S., and K. D. Entian. Function of Lactococcus lactis Nisin Immunity Genes nisI and nisFEG after Coordinated Expression in the Surrogate Host Bacillus subtilis. The Jurnal of Biological Chemistry, USA. , Vol. 278, pp. 89 -94 (2003)
[2] Yu-Kuo Tsai, Hung- Wen Chen, Ta-Chun Lo and Thy-Hou Lin. Specific point mutation in Lactobacillus casei ATCC 27139 cause the phenotype switch from Lac- to Lac+. Microbiology, pp. 751-760(2009)
[3] Ju-Chen Cheng, 2009, Investigation on the Characteristic of Heparin-Binding Haemagglutinin 3 Adhesin (HBHA)-Related Peptides and their Transcytosis
[4] C. W. le Roux and S. R. Bloom. Peptide YY, appetite and food intake.(2005)
Maintained by the iGEM team NTU-LIHPAO-Taiwan    ©2015 NTU-LIHPAO-Taiwan