Difference between revisions of "Part:BBa K1890000"
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<h2>Introduction</h2>
 
<h2>Introduction</h2>
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Silicatein, originating from the demosponge <i>Suberites domuncula</i>, catalyzes the formation of polysilicate. This biobrick contains the short version of the silicatein gene, according to Müller <i>et al</i> [1][2]. The silicatein gene is combined with the strong RBS <partinfo>BBa_B0034</partinfo>.
 
Silicatein, originating from the demosponge <i>Suberites domuncula</i>, catalyzes the formation of polysilicate. This biobrick contains the short version of the silicatein gene, according to Müller <i>et al</i> [1][2]. The silicatein gene is combined with the strong RBS <partinfo>BBa_B0034</partinfo>.
  
 
<h2>Sequence and Features</h2>
 
<h2>Sequence and Features</h2>
 
<partinfo>BBa_K1890000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1890000 SequenceAndFeatures</partinfo>
 +
 +
<h2>Usage and Biology</h2>
 +
Silicatein is an enzyme natively found in demosponges and diatoms,
 +
where it catalyzes the condensation of silica to form the typical skeletal elements.
 +
Here, we use the enzyme to create a polysilicate layer around the host organism <i>E. coli</i>.
 +
 +
<h2>Characterization</h2>
 +
In order to characterize the formation of a polysilicate layer around <i>E. coli</i>, we performed the following experiments.
 +
<ul><li>Rhodamine 123 staining</li>
 +
    <li> Growth study</li></ul>
 +
 +
<h3>Staining with Rhodamine 123</h3>
 +
In this experiment we imaged the silicatein expressing cells with a fluorescence microscope, after treating them with a fluorescent dye.
 +
The fluorescent dye Rhodamine 123 (Sigma) has shown to bind specifically to polysilicate [3].
 +
Cells were stained according to the protocol based on Li <i>et al.</i> and Müller <i>et al.</i> [3][4].
 +
Rhodamine was excited with a wavelength of 395 nm.
  
 
<h2>References</h2>
 
<h2>References</h2>
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[2] Müller, W. E. G. (2003). Silicon biomineralization.
 
[2] Müller, W. E. G. (2003). Silicon biomineralization.
 +
 +
[3] Li, C. W., Chu, S., & Lee, M. (1989). Characterizing the silica deposition vesicle of diatoms. Protoplasma, 151(2-3), 158–163.
 +
 +
[4] Müller, W. E. G., Rothenberger, M., Boreiko, A., Tremel, W., Reiber, A., & Schröder, H. C. (2005). Formation of siliceous spicules in the marine demosponge Suberites domuncula. Cell and Tissue Research, 321(2), 285–297.

Revision as of 18:49, 17 October 2016

Silicatein gene with strong RBS

Introduction

Silicatein, originating from the demosponge Suberites domuncula, catalyzes the formation of polysilicate. This biobrick contains the short version of the silicatein gene, according to Müller et al [1][2]. The silicatein gene is combined with the strong RBS BBa_B0034.

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]

Usage and Biology

Silicatein is an enzyme natively found in demosponges and diatoms, where it catalyzes the condensation of silica to form the typical skeletal elements. Here, we use the enzyme to create a polysilicate layer around the host organism E. coli.

Characterization

In order to characterize the formation of a polysilicate layer around E. coli, we performed the following experiments.

  • Rhodamine 123 staining
  • Growth study

Staining with Rhodamine 123

In this experiment we imaged the silicatein expressing cells with a fluorescence microscope, after treating them with a fluorescent dye. The fluorescent dye Rhodamine 123 (Sigma) has shown to bind specifically to polysilicate [3]. Cells were stained according to the protocol based on Li et al. and Müller et al. [3][4]. Rhodamine was excited with a wavelength of 395 nm.

References

[1] Müller, W. E. G., Engel, S., Wang, X., Wolf, S. E., Tremel, W., Thakur, N. L., … Schröder, H. C. (2008). Bioencapsulation of living bacteria (Escherichia coli) with poly(silicate) after transformation with silicatein-α gene. Biomaterials, 29(7), 771–779.

[2] Müller, W. E. G. (2003). Silicon biomineralization.

[3] Li, C. W., Chu, S., & Lee, M. (1989). Characterizing the silica deposition vesicle of diatoms. Protoplasma, 151(2-3), 158–163.

[4] Müller, W. E. G., Rothenberger, M., Boreiko, A., Tremel, W., Reiber, A., & Schröder, H. C. (2005). Formation of siliceous spicules in the marine demosponge Suberites domuncula. Cell and Tissue Research, 321(2), 285–297.