Team:Tsinghua/BioBricks

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<h1>Biobricks</h1>
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<p>
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Pathogen detection is one major topic related to the access to health care, the failing of which leads to serious consequences. Pseudomonasaeruginosa and Staphylococcus aureusare two problematic pathogenic Gram-negative bacteria causing various diseases.Fast and sensitive detection of them is required for rapidly administered andappropriate antibiotic treatment in serious medical conditions.
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</p>
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By the inspiration of quorum sensing system, we designed a novel pathogen detection system.  Considering the specificity and sensitivity of quorum sensing, this technology could prove useful forclinical test, medical diagnostics, and other potential applications.
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</p>
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<li class="section1">
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BBa_K1024000
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<li class="section2">
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BBa_K1024001
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<li class="section3">
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BBa_K1024002
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<li class="section4">
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BBa_K1024003
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<div class="section section1">
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<h2>BBa_K1024000</h2>
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<div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/a/a9/Tsinghua-Biobrick1.png" width="100%"/>
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<p>
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<b>Name:</b> BBa_K1024000
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</p>
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<p>
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<b>Short Description:</b> LuxR in Yeast (pTEF+VP16+NLS+LuxR)
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</p>
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<p>
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<b>Part Type:</b> Regulatory
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</p>
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<p>
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<b>Design:</b> We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). We modified the transcription activator, LuxR (BBa_C0062), by adding nuclear localization signal (NLS) sequence and Herpes simplex virus VP16 activation domain in N-terminus of LuxR, and ligate the sequence of this modified LuxR downstream TEF promoter, which is the constitutive promoter in yeast (BBa_K1024000).
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</p>
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<p>
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<b>Reference:</b>
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Fuqua W C, Winans S C, Greenberg E P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators[J]. Journal of bacteriology, 1994, 176(2): 269.
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Sadowski I, Ma J, Triezenberg S, et al. GAL4-VP16 is an unusually potent transcriptional activator[J]. Nature, 1988, 335(6190): 563-564.
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</p>
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<div class="section section2">
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<h2>BBa_K1024001</h2>
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<div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/8/80/Tsinghua-Biobrick2.png" width="100%"/>
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</div>
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<p>
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<b>Part Name:</b> BBa_K1024001
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</p>
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<p>
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<b>Short Description:</b> Lux Promoter in Yeast (pLux+Cyc Promoter+mCherry)
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</p>
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<p>
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<b>Part Type:</b> Reporter
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</p>
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<p>
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<b>Design:</b> We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). We modified the transcriptional regulated promoter in quorum sensing system, Plux promoter (BBa_R0062), by adding cyc100 mini promoter downstream of the Plux promoter (BBa_K1024001).
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</p>
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<p>
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<b>Reference:</b>
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Fuqua W C, Winans S C, Greenberg E P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators[J]. Journal of bacteriology, 1994, 176(2): 269.
 +
Sadowski I, Ma J, Triezenberg S, et al. GAL4-VP16 is an unusually potent transcriptional activator[J]. Nature, 1988, 335(6190): 563-564.
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</p>
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</div>
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<div class="section section3">
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<h2>BBa_K1024002</h2>
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<div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/9/94/Tsinghua-Biobrick3.png" width="100%"/>
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</div>
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<p>
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<b>Part Name:</b> BBa_K1024002
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</p>
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<p>
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<b>Short Description:</b> Reporter for quorum sensing systems in yeast
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</p>
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<p>
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<b>Part Type:</b> Signaling
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</p>
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<p>
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<b>Design:</b> We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). The LuxR gene is constitutively expressed, while the activation of Lux Promoter requires the signaling of N-Acyl Homoserine Lactone (AHL). Therefore, mCherry is activated in the presence of AHL.
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</p>
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<p>
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<b>Test:</b> <div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/f/f2/Tsinghua-Biobrick4.png" width="100%"/>
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</div>
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</p>
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<p>
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<b>Reference:</b>
 +
Fuqua W C, Winans S C, Greenberg E P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators[J]. Journal of bacteriology, 1994, 176(2): 269.
 +
Sadowski I, Ma J, Triezenberg S, et al. GAL4-VP16 is an unusually potent transcriptional activator[J]. Nature, 1988, 335(6190): 563-564.
 +
 
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</p>
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</div>
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<div class="section section4">
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<h2>BBa_K1024003</h2>
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<div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/2/23/Tsinghua-Biobrick5.png" width="100%"/>
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</div>
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<p>
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<b>Part Name:</b> BBa_K1024003
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</p>
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<p>
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<b>Short Description:</b> Reporter for Tet system in Yeast
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</p>
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<p>
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<b>Part Type:</b> Signaling
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</p>
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<p>
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<b>Design:</b> The part is designed to valid the Tet system in yeast. TetR gene with VP16 is constitutively expressed, activating the downstream reporter gene of TetO and CYC1 TATA region. In the yeast Saccharomyces cerevisiae the ade2, and/or the ade1, mutation in the adenine biosynthetic pathway leads to the accumulation of a cell-limited red pigment. Thus, it could be used as a marker for screening of target phenotype.
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<div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/f/f2/Tsinghua-Biobrick4.png" width="100%"/>
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</div>
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</p>
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<p>
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<b>Test:</b> Yeast with ADE2 knockout exhibits red in color. The function of the plasmid was tested by rescuing the knockout strains, which regained the white color by expressing ADE2.
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<div class="right" style="width: 400px">
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<img height="auto" src="https://static.igem.org/mediawiki/2013/1/19/Tsinghua-Biobrick7_%282%29.JPG" width="100%"/>
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</div>
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</p>
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<p>
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<b>Reference:</b>
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Bellí G, Garí E, Piedrafita L, et al. An activator/repressor dual system allows tight tetracycline-regulated gene expression in budding yeast[J]. Nucleic acids research, 1998, 26(4): 942-947.
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Revision as of 11:34, 27 September 2013

Biobricks

Pathogen detection is one major topic related to the access to health care, the failing of which leads to serious consequences. Pseudomonasaeruginosa and Staphylococcus aureusare two problematic pathogenic Gram-negative bacteria causing various diseases.Fast and sensitive detection of them is required for rapidly administered andappropriate antibiotic treatment in serious medical conditions.

By the inspiration of quorum sensing system, we designed a novel pathogen detection system.  Considering the specificity and sensitivity of quorum sensing, this technology could prove useful forclinical test, medical diagnostics, and other potential applications.

  • BBa_K1024000
  • BBa_K1024001
  • BBa_K1024002
  • BBa_K1024003

BBa_K1024000

Name: BBa_K1024000

Short Description: LuxR in Yeast (pTEF+VP16+NLS+LuxR)

Part Type: Regulatory

Design: We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). We modified the transcription activator, LuxR (BBa_C0062), by adding nuclear localization signal (NLS) sequence and Herpes simplex virus VP16 activation domain in N-terminus of LuxR, and ligate the sequence of this modified LuxR downstream TEF promoter, which is the constitutive promoter in yeast (BBa_K1024000).

Reference: Fuqua W C, Winans S C, Greenberg E P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators[J]. Journal of bacteriology, 1994, 176(2): 269. Sadowski I, Ma J, Triezenberg S, et al. GAL4-VP16 is an unusually potent transcriptional activator[J]. Nature, 1988, 335(6190): 563-564.

BBa_K1024001

Part Name: BBa_K1024001

Short Description: Lux Promoter in Yeast (pLux+Cyc Promoter+mCherry)

Part Type: Reporter

Design: We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). We modified the transcriptional regulated promoter in quorum sensing system, Plux promoter (BBa_R0062), by adding cyc100 mini promoter downstream of the Plux promoter (BBa_K1024001).

Reference: Fuqua W C, Winans S C, Greenberg E P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators[J]. Journal of bacteriology, 1994, 176(2): 269. Sadowski I, Ma J, Triezenberg S, et al. GAL4-VP16 is an unusually potent transcriptional activator[J]. Nature, 1988, 335(6190): 563-564.

BBa_K1024002

Part Name: BBa_K1024002

Short Description: Reporter for quorum sensing systems in yeast

Part Type: Signaling

Design: We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). The LuxR gene is constitutively expressed, while the activation of Lux Promoter requires the signaling of N-Acyl Homoserine Lactone (AHL). Therefore, mCherry is activated in the presence of AHL.

Test:

Reference: Fuqua W C, Winans S C, Greenberg E P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators[J]. Journal of bacteriology, 1994, 176(2): 269. Sadowski I, Ma J, Triezenberg S, et al. GAL4-VP16 is an unusually potent transcriptional activator[J]. Nature, 1988, 335(6190): 563-564.

BBa_K1024003

Part Name: BBa_K1024003

Short Description: Reporter for Tet system in Yeast

Part Type: Signaling

Design: The part is designed to valid the Tet system in yeast. TetR gene with VP16 is constitutively expressed, activating the downstream reporter gene of TetO and CYC1 TATA region. In the yeast Saccharomyces cerevisiae the ade2, and/or the ade1, mutation in the adenine biosynthetic pathway leads to the accumulation of a cell-limited red pigment. Thus, it could be used as a marker for screening of target phenotype.

Test: Yeast with ADE2 knockout exhibits red in color. The function of the plasmid was tested by rescuing the knockout strains, which regained the white color by expressing ADE2.

Reference: Bellí G, Garí E, Piedrafita L, et al. An activator/repressor dual system allows tight tetracycline-regulated gene expression in budding yeast[J]. Nucleic acids research, 1998, 26(4): 942-947.

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