Difference between revisions of "Part:BBa K1724001"
 
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<partinfo>BBa_K1724001 short</partinfo>
 
<partinfo>BBa_K1724001 short</partinfo>
  
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The mercury –sensing regulatory protein, MerR(wild type), which regulates mercury resistance operons in Gram-negative bacteria, is subjected to directed evolution in an effect to generate a MerR mutant that responds to Cadmium ion but not mercury.That is, the MerR mutant is the cadmium-sensing regulatory protein. To get MerR mutant, Oligonucleotide-directed mutagenesis is used to introduce random mutations into the key metal-binding regions of MerR. Finally, Getting the generated Cd-specific MerR mutants appears to be unique.
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Curli, the first identified functional amyloid fibres, are extracellular protein fibers produced by many enteric bacteria including Escherichia coli and Salmonella species [1]. The curli system exhibits numerous features that make it an ideal platform for the type of materials engineering by way of synthetic biology that we envision. First, as the curli nanofibre is composed primarily from the self-assembly of one small protein, it presents a tractable entry point towards creating a large diversity of biofilm extracellular matrices with conventional genetic engineering methods. Second, the functional amyloid fibres formed by CsgA are extremely robust, being able to withstand boiling in detergents and extended incubation in solvents, increasing their potential utility in harsh environments. Finally, recent findings have shown that the curli system can be used to efficiently export natively unfolded polypeptides and can be used in a broad and modular way for the display of various functional peptides throughout the E. coli biofilm [2]. CsgA, the dominant proteinaceous component in E. coli biofilms, is capable of self-polymerizing in vitro into β-sheet-rich amyloid fibers that bind to the amyloid specific dye Congo red (CR), resulting in a red shift and green birefringence under polarized light, and thioflavin T (ThT), leading to increased fluorescence at certain wavelengths [3].
  
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The quantification data of MerR/CadA operon can be seen in BBa_K1724000.[[https://parts.igem.org/Part:BBa_K1724000]]
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== Reference ==
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[1] Evans, M.L., and Chapman, M.R. (2014). Curli biogenesis: order out of disorder. Biochimica et biophysica acta 1843, 1551-1558.
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[2] Nguyen, P.Q., Botyanszki, Z., Tay, P.K., and Joshi, N.S. (2014). Programmable biofilm-based materials from engineered curli nanofibres. Nature communications 5, 4945.
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[3] Blanco, L.P., Evans, M.L., Smith, D.R., Badtke, M.P., and Chapman, M.R. (2012). Diversity, biogenesis and function of microbial amyloids. Trends in microbiology 20, 66-73.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 08:06, 15 September 2015

RBS+CsgA

Curli, the first identified functional amyloid fibres, are extracellular protein fibers produced by many enteric bacteria including Escherichia coli and Salmonella species [1]. The curli system exhibits numerous features that make it an ideal platform for the type of materials engineering by way of synthetic biology that we envision. First, as the curli nanofibre is composed primarily from the self-assembly of one small protein, it presents a tractable entry point towards creating a large diversity of biofilm extracellular matrices with conventional genetic engineering methods. Second, the functional amyloid fibres formed by CsgA are extremely robust, being able to withstand boiling in detergents and extended incubation in solvents, increasing their potential utility in harsh environments. Finally, recent findings have shown that the curli system can be used to efficiently export natively unfolded polypeptides and can be used in a broad and modular way for the display of various functional peptides throughout the E. coli biofilm [2]. CsgA, the dominant proteinaceous component in E. coli biofilms, is capable of self-polymerizing in vitro into β-sheet-rich amyloid fibers that bind to the amyloid specific dye Congo red (CR), resulting in a red shift and green birefringence under polarized light, and thioflavin T (ThT), leading to increased fluorescence at certain wavelengths [3].


Reference

[1] Evans, M.L., and Chapman, M.R. (2014). Curli biogenesis: order out of disorder. Biochimica et biophysica acta 1843, 1551-1558.

[2] Nguyen, P.Q., Botyanszki, Z., Tay, P.K., and Joshi, N.S. (2014). Programmable biofilm-based materials from engineered curli nanofibres. Nature communications 5, 4945.

[3] Blanco, L.P., Evans, M.L., Smith, D.R., Badtke, M.P., and Chapman, M.R. (2012). Diversity, biogenesis and function of microbial amyloids. Trends in microbiology 20, 66-73.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]