Difference between revisions of "Part:BBa K2924026"
 
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The DNA sequence of the gene was acquired by reverse-translating the amino acid sequence. Further, in order to reach optimal heterologous expression in <i>Escherichia coli</i> the DNA sequence was designed with optimized codons and synthesized commercially.
 
The DNA sequence of the gene was acquired by reverse-translating the amino acid sequence. Further, in order to reach optimal heterologous expression in <i>Escherichia coli</i> the DNA sequence was designed with optimized codons and synthesized commercially.
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<h2>Fire Retardancy Test</h2>
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<p> The test subject(Figure 1.) was in the size of bed sheet in 300*130mm and the average Moisture content% is 13.55%-14.37%. As demonstrated on Fig. 1, all of our engineered flame retardant displayed a significant improvement compared with the water control.</p>
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<img src="https://2020.igem.org/wiki/images/9/9c/T--Puiching_Macau--bedsheet_chart.png">
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<h2>Contribution</h2>
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<p>Team: PuiChing_Macau 2020<br>
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Summary: We have compared the original alpha-casein without adhesion domains and our engineered alpha casein with adhesion domains, by the fire retardancy test and added the RFP to the alpha casein in order to observe the adhesive test result under fluorescents microscope. As shown in Figures 1 & 2, the protein with adhesion domain has a great improvement on its fire retardancy. In this figure, the results show a clear improvement of the adhesiveness of the fire retardant protein.
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<img src="https://2020.igem.org/wiki/images/9/9c/T--Puiching_Macau--bedsheet_chart.png" width="600">
 
<p> Figure 1. Vertical Burning Test using bedsheet</p>
 
<p> Figure 1. Vertical Burning Test using bedsheet</p>
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<p>  The test object was a wooden block with the size of 40*38*51mm and the average Moisture content% is 12.6%. All of the engineered  flame retardants displayed a significant improvement compared with the water control. </p>
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<img src="https://2020.igem.org/wiki/images/2/2e/T--Puiching_Macau--wood.png">
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<img src="https://2020.igem.org/wiki/images/2/2e/T--Puiching_Macau--wood.png" width="600">
 
<p> Figure 2. Fire retardant test for wood</p>
 
<p> Figure 2. Fire retardant test for wood</p>
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<h2>Adhesion test</h2>
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<h4>Adhesion test</h4>
 
<p>  Figure 3. The adhesion level of (3b)K3503009(RFP-Alpha Casein), (3c)K3503011(RFP-mfp5-Alpha casein),(3d)K3503010 (RFP-CBD-Alpha casein)  by different state of process(non- washing,  after washing with water and soaking.) </p>
 
<p>  Figure 3. The adhesion level of (3b)K3503009(RFP-Alpha Casein), (3c)K3503011(RFP-mfp5-Alpha casein),(3d)K3503010 (RFP-CBD-Alpha casein)  by different state of process(non- washing,  after washing with water and soaking.) </p>
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<img src="https://2020.igem.org/wiki/images/6/66/T--Puiching_Macau--The_percentage.png">
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<img src="https://2020.igem.org/wiki/images/6/66/T--Puiching_Macau--The_percentage.png" width="600">
 
<p> Figure 3(a) The percentage of the protein after processes</p>
 
<p> Figure 3(a) The percentage of the protein after processes</p>
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<img src="https://2020.igem.org/wiki/images/7/7a/T--Puiching_Macau--alpha_RFP.png">
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<img src="https://2020.igem.org/wiki/images/7/7a/T--Puiching_Macau--alpha_RFP.png" width="600">
 
<p> Figure 3(b) K3503009(RFP-Alpha Casein)</p>
 
<p> Figure 3(b) K3503009(RFP-Alpha Casein)</p>
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<img src="https://2020.igem.org/wiki/images/0/04/T--Puiching_Macau--mfp5_RFP.png">
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<img src="https://2020.igem.org/wiki/images/0/04/T--Puiching_Macau--mfp5_RFP.png" width="600">
 
<p> Figure 3(c) K3503011(RFP-mfp5-Alpha casein)</p>
 
<p> Figure 3(c) K3503011(RFP-mfp5-Alpha casein)</p>
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<img src="https://2020.igem.org/wiki/images/f/f1/T--Puiching_Macau--CBD_RFP.png">
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<img src="https://2020.igem.org/wiki/images/f/fb/T--Puiching_Macau--RFP-CBD.png" width="600">
 
<p> Figure 3(d) K3503010 (RFP-CBD-Alpha casein) </p>
 
<p> Figure 3(d) K3503010 (RFP-CBD-Alpha casein) </p>
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<p>Figure 3.The adhesion level of (3b)K3503009(RFP-Alpha Casein), (3c)K3503011(RFP-mfp5-Alpha casein),(3d)K3503010 (RFP-CBD-Alpha casein)  by different state of process(non- washing,  after washing with water and soaking.)</p>
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<h4>Conclusion</h4>
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<p> Overall, the proteins with adhesion retain a higher fire retardancy than the protein without adhesion through adhesive test and fire retardancy test. Therefore, the genetic fusion between alpha casein and adhesion domain would generate a more stable fire retardant protein.</p>
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Latest revision as of 22:16, 27 October 2020


α-s1-casein

The milk protein α-s1-casein from Bos taurus

Usage and Biology

α-s1-casein (CSN1S1), also named B5B3R8_BOVIN1is a gene coding for a protein product and has the GeneID: 282208, which originates from 2.

Caseins are among the most abundant proteins in native cow's milk. They are phosphoproteins and make up approximately 80% of total dairy protein mass. The milk proteins and their proportions are important to the parameters of milk manufacturing and the quality of milk protein. With a length of 17508 bp and 19 exons the bovine α-s1-casein gene is the most abundant one of the bovine caseins (12-15 g/L)3. The α-s1-casein protein may contain up to eight serine monophosphate residues, which cluster in a hydrophilic domain between amino acids 43–80 and, through modelling studies, are thought to be connected to a hydrophobic domain (amino acids 100–199) by helical and sheet secondary structures4. In the ability of milk to carry calcium phosphate, α-s1-casein also performs an significant part5.

The DNA sequence of the gene was acquired by reverse-translating the amino acid sequence. Further, in order to reach optimal heterologous expression in Escherichia coli the DNA sequence was designed with optimized codons and synthesized commercially.

Contribution

Team: PuiChing_Macau 2020
Summary: We have compared the original alpha-casein without adhesion domains and our engineered alpha casein with adhesion domains, by the fire retardancy test and added the RFP to the alpha casein in order to observe the adhesive test result under fluorescents microscope. As shown in Figures 1 & 2, the protein with adhesion domain has a great improvement on its fire retardancy. In this figure, the results show a clear improvement of the adhesiveness of the fire retardant protein.

Figure 1. Vertical Burning Test using bedsheet

Figure 2. Fire retardant test for wood

Adhesion test

Figure 3. The adhesion level of (3b)K3503009(RFP-Alpha Casein), (3c)K3503011(RFP-mfp5-Alpha casein),(3d)K3503010 (RFP-CBD-Alpha casein) by different state of process(non- washing, after washing with water and soaking.)

Figure 3(a) The percentage of the protein after processes

Figure 3(b) K3503009(RFP-Alpha Casein)

Figure 3(c) K3503011(RFP-mfp5-Alpha casein)

Figure 3(d) K3503010 (RFP-CBD-Alpha casein)

Figure 3.The adhesion level of (3b)K3503009(RFP-Alpha Casein), (3c)K3503011(RFP-mfp5-Alpha casein),(3d)K3503010 (RFP-CBD-Alpha casein) by different state of process(non- washing, after washing with water and soaking.)

Conclusion

Overall, the proteins with adhesion retain a higher fire retardancy than the protein without adhesion through adhesive test and fire retardancy test. Therefore, the genetic fusion between alpha casein and adhesion domain would generate a more stable fire retardant protein.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 643
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1]: https://www.uniprot.org/uniprot/B5B3R8

[2]: https://www.ncbi.nlm.nih.gov/gene/?term=282208

[3]: Boland, Mike, Harjinder Singh, and Abby Thompson, eds. Milk proteins: from expression to food. Academic Press, 2014.

[4]: Sheehy, P. A., et al. "Significance, origin and function of bovine milk proteins: the biological implications of manipulation or modification." Milk Proteins. Academic Press, 2008. 81-106.

[5]: Chanat, E., J. Cell Sci., 112, 3399-3412 (1999)