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Team:Tsinghua/BioBricks
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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. | <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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<img height="auto" src="https://static.igem.org/mediawiki/2013/f/f2/Tsinghua-Biobrick4.png" width="100%"/> | <img height="auto" src="https://static.igem.org/mediawiki/2013/f/f2/Tsinghua-Biobrick4.png" width="100%"/> | ||
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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. | <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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<img height="auto" src="https://static.igem.org/mediawiki/2013/1/19/Tsinghua-Biobrick7_%282%29.JPG" width="100%"/> | <img height="auto" src="https://static.igem.org/mediawiki/2013/1/19/Tsinghua-Biobrick7_%282%29.JPG" width="100%"/> | ||
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<b>Reference:</b> | <b>Reference:</b> | ||
Revision as of 11:42, 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
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.
