Difference between revisions of "Part:BBa K4814001"
 
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<partinfo>BBa_K4814001 short</partinfo>
 
<partinfo>BBa_K4814001 short</partinfo>
  
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<h1>RecA(K6)-hha</h1>
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This is a composite part aimed to kill the bacteria after testing for DNA damage. The RecA promoter is BBa_K629001, followed by RBS (BBa_B0034), biofilm reducing protein hha (BBa_K484000), and terminator (BBa_B0015). Experiments and data shows that by using hha, after the treatment of carcinogens, the optical density of the sample decreases more significantly than the control group (E.coli top10). Detailed information of hha can be obtained from BBa_K4814000.
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<p><span style="color: rgb(0, 0, 0);font-size: 12pt;">This is a composite part aimed to kill the bacteria after testing for DNA damage. The RecA promoter is BBa_K629001, followed by RBS (BBa_B0034), biofilm reducing protein hha (BBa_K484000), and terminator (BBa_B0015). Experiments and data shows that by using hha, after the treatment of carcinogens, the optical density of the sample decreases more significantly than the control group (E.coli top10). Detailed information of hha can be obtained from BBa_K4814000.</span></p>
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<p><span style="color: rgb(0, 0, 0);font-size: 12pt;">BBa_K629001 [ccaataattttgt] BBa_B0034 [atatacat] BBa_K4814000 [cgcggatccgg] BBa_B0015</span></p>
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Initially, we hypothesized that as the concentration of carcinogens increased, the OD would decrease due to the bactericidal effects of the activated hha protein. However, the experimental results revealed that despite the escalating level of carcinogen treatment, the OD did not exhibit a significant decrease compared to the control group (top10 or E. coli), nor did it demonstrate a discernible trend. This could be attributed to adaptive responses to stress or incomplete protein folding of the hha protein.
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As illustrated in Figure 1, the OD of the TOP10 control group decreased slightly from 0.28 to 0.26, and then increased to 0.3 after 6, 12, and 18 minutes of UVB exposure. In contrast, starting at 0.24, the OD of the hha group showed a slight increase and then decreased to 0.22 after 18 minutes of UVB treatment. In the graph representing the H2O2 group (Figure 2), both the hha and TOP10 groups exhibited a decreasing trend in OD, consistent with our initial hypothesis.
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To further investigate this phenomenon, future studies can assess the expression level of the hha protein using techniques such as Western Blot or qPCR. These analyses will provide insights into the protein's expression patterns and help elucidate the underlying mechanisms contributing to the observed OD changes.
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<html>
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<table>
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<tr>
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<td><img src=https://static.igem.wiki/teams/4814/wiki/lab/hha-uvb.png alt=""style="width:400px;height:auto;">
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</td>
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<td>
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<img src=https://static.igem.wiki/teams/4814/wiki/lab/hha-h2o2.png alt=""style="width:400px;height:auto;">
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</td>
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</tr>
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</table>
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<p>Figure 1 & 2. The graph of the OD600 of RecA(K6)-hha after being treated with UVB (left) and H2O2 (right).</p>
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<table>
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<tr>
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<td>
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<img src=https://static.igem.wiki/teams/4814/wiki/lab/hha-na.png alt=""style="width:400px;height:auto;">
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</td>
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<td>
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<img src=https://static.igem.wiki/teams/4814/wiki/lab/hha-ap.png alt=""style="width:400px;height:auto;">
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</td>
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</tr>
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</table>
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<p>Figure 3 & 4. The graph of the OD600 of RecA(K6)-hha after being treated with Nalidixic acid (left) and Aspartame (right).</p>
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</html>
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References:
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Balsalobre, C., Juárez, A., Madrid, C., Mouriño, M., Prenafeta, A., & Muñoa, F. J. (1996). Complementation of the hha mutation in Escherichia coli by the ymoA gene from Yersinia enterocolitica: dependence on the gene dosage. Microbiology (Reading, England), 142 ( Pt 7), 1841–1846. https://doi.org/10.1099/13500872-142-7-1841
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Hong, S. H., Lee, J., & Wood, T. K. (2010). Engineering global regulator Hha of Escherichia coli to control biofilm dispersal. Microbial biotechnology, 3(6), 717–728. https://doi.org/10.1111/j.1751-7915.2010.00220.x
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Madrid, C., Nieto, J. M., & Juárez, A. (2001). Role of the Hha/YmoA family of proteins in the thermoregulation of the expression of virulence factors. International Journal of Medical Microbiology, 291(6-7), 425-432. https://doi.org/10.1078/1438-4221-00149
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Latest revision as of 04:52, 10 October 2023


RecA(K6)-hha

This is a composite part aimed to kill the bacteria after testing for DNA damage. The RecA promoter is BBa_K629001, followed by RBS (BBa_B0034), biofilm reducing protein hha (BBa_K484000), and terminator (BBa_B0015). Experiments and data shows that by using hha, after the treatment of carcinogens, the optical density of the sample decreases more significantly than the control group (E.coli top10). Detailed information of hha can be obtained from BBa_K4814000.

Initially, we hypothesized that as the concentration of carcinogens increased, the OD would decrease due to the bactericidal effects of the activated hha protein. However, the experimental results revealed that despite the escalating level of carcinogen treatment, the OD did not exhibit a significant decrease compared to the control group (top10 or E. coli), nor did it demonstrate a discernible trend. This could be attributed to adaptive responses to stress or incomplete protein folding of the hha protein.

As illustrated in Figure 1, the OD of the TOP10 control group decreased slightly from 0.28 to 0.26, and then increased to 0.3 after 6, 12, and 18 minutes of UVB exposure. In contrast, starting at 0.24, the OD of the hha group showed a slight increase and then decreased to 0.22 after 18 minutes of UVB treatment. In the graph representing the H2O2 group (Figure 2), both the hha and TOP10 groups exhibited a decreasing trend in OD, consistent with our initial hypothesis.

To further investigate this phenomenon, future studies can assess the expression level of the hha protein using techniques such as Western Blot or qPCR. These analyses will provide insights into the protein's expression patterns and help elucidate the underlying mechanisms contributing to the observed OD changes.

Figure 1 & 2. The graph of the OD600 of RecA(K6)-hha after being treated with UVB (left) and H2O2 (right).

Figure 3 & 4. The graph of the OD600 of RecA(K6)-hha after being treated with Nalidixic acid (left) and Aspartame (right).


References:

Balsalobre, C., Juárez, A., Madrid, C., Mouriño, M., Prenafeta, A., & Muñoa, F. J. (1996). Complementation of the hha mutation in Escherichia coli by the ymoA gene from Yersinia enterocolitica: dependence on the gene dosage. Microbiology (Reading, England), 142 ( Pt 7), 1841–1846. https://doi.org/10.1099/13500872-142-7-1841

Hong, S. H., Lee, J., & Wood, T. K. (2010). Engineering global regulator Hha of Escherichia coli to control biofilm dispersal. Microbial biotechnology, 3(6), 717–728. https://doi.org/10.1111/j.1751-7915.2010.00220.x

Madrid, C., Nieto, J. M., & Juárez, A. (2001). Role of the Hha/YmoA family of proteins in the thermoregulation of the expression of virulence factors. International Journal of Medical Microbiology, 291(6-7), 425-432. https://doi.org/10.1078/1438-4221-00149


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 383
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]