Difference between revisions of "Part:BBa K1949052"
 
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<partinfo>BBa_K1949052 short</partinfo>
 
<partinfo>BBa_K1949052 short</partinfo>
  
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<span style="margin-left: 10px;">amiE codes for protein AmiE. AmiE is an acylase that degrades long chain N-acyl homoserine lactone (AHL) molecules with acyl chains longer than eightsix carbons.
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<span style="margin-left: 10px;">This gene codes for protein AmiE. AmiE is an acylase that degrades long chain N-acyl homoserine lactone (AHL) molecules with acyl chains longer than eightsix carbons.
  
  
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Our objective was to characterize the function of AmiE protein. We prepared three samples as shown below. When we tested the AmiE degradation ability with these samples, the results show that C4HSL is not degraded by AmiE, but 3OC12HSL is degraded by AmiE.
 
Our objective was to characterize the function of AmiE protein. We prepared three samples as shown below. When we tested the AmiE degradation ability with these samples, the results show that C4HSL is not degraded by AmiE, but 3OC12HSL is degraded by AmiE.
  
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・PBAD/araC-rbs-AmiE(pSB6A1)
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・PBAD/araC-<i>rbs-AmiE</i>(pSB6A1)
  
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・Ptet-rbs-luxR-TT-Plux-rbs-gfp (pSB6A1)
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・Ptet-<i>rbs-lux</i>R-<i>tt</i>-Plux-<i>rbs-gfp</i> (pSB6A1)
  
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・Ptet-rbs-luxR-TT-Plux-rbs-gfp (pSB6A1)
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・Ptet-<i>rbs-rhl</i>R-<i>tt</i>-Plux-<i>rbs-gfp</i> (pSB6A1)
  
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[[Image:amiE2.png|thumb|center|400px| Fig.1 Incase of C4HSL, the fluorescence intensity of the reporter with the supernatant solution induced the AmiE expression was almost the same as the one with the supernatant solution not induced the AmiE expression. ]]<br>
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[[Image:amie1.png|thumb|center|400px| Fig.1 Incase of C4HSL, the fluorescence intensity of the reporter with the supernatant solution induced the AmiE expression was almost the same as the one with the supernatant solution not induced the AmiE expression. ]]<br>
  
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[[Image:amiE3.png|thumb|center|400px| Fig.2 In case of 3OC12HSL, the fluorescence intensity of the reporter added the solution induced the AmiE expression was about 1/10 of the reporter added the solution not induced AmiE expression. ]]<br>
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[[Image:amie2.png|thumb|center|400px| Fig.2 In case of 3OC12HSL, the fluorescence intensity of the reporter added the solution induced the AmiE expression was about 1/10 of the reporter added the solution not induced AmiE expression. ]]<br>
  
 
<span style="margin-left: 10px;">This graph shows that the fluorescence measurement of the reporters added the E. coli solution both induced AmiE expression and not.
 
<span style="margin-left: 10px;">This graph shows that the fluorescence measurement of the reporters added the E. coli solution both induced AmiE expression and not.
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However, in case of 3OC12HSL, the fluorescence intensity of the reporter added the solution induced the AmiE expression was about 1/10 of the reporter added the solution not induced AmiE expression. Based on the above, it is concluded that AmiE does not degrade C4HSL but degrades 3OC12HSL.(Fig.2)
 
However, in case of 3OC12HSL, the fluorescence intensity of the reporter added the solution induced the AmiE expression was about 1/10 of the reporter added the solution not induced AmiE expression. Based on the above, it is concluded that AmiE does not degrade C4HSL but degrades 3OC12HSL.(Fig.2)
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Latest revision as of 20:27, 18 October 2016


PBAD-rbs-amiE

This gene codes for protein AmiE. AmiE is an acylase that degrades long chain N-acyl homoserine lactone (AHL) molecules with acyl chains longer than eightsix carbons.


Fig.1 AmiE is an acylase that degrades long chain N-acyl homoserine lactone (AHL) molecules with acyl chains longer than eightsix carbons

Characterization

We tested the function of AmiE protein that influences the end of the story.

Our objective was to characterize the function of AmiE protein. We prepared three samples as shown below. When we tested the AmiE degradation ability with these samples, the results show that C4HSL is not degraded by AmiE, but 3OC12HSL is degraded by AmiE.

・PBAD/araC-rbs-AmiE(pSB6A1)

・Ptet-rbs-luxR-tt-Plux-rbs-gfp (pSB6A1)

・Ptet-rbs-rhlR-tt-Plux-rbs-gfp (pSB6A1)

Fig.1 Incase of C4HSL, the fluorescence intensity of the reporter with the supernatant solution induced the AmiE expression was almost the same as the one with the supernatant solution not induced the AmiE expression.

Fig.2 In case of 3OC12HSL, the fluorescence intensity of the reporter added the solution induced the AmiE expression was about 1/10 of the reporter added the solution not induced AmiE expression.

This graph shows that the fluorescence measurement of the reporters added the E. coli solution both induced AmiE expression and not.

From the results of the AmiE degradation activity toward C4HSL, the fluorescence intensity of the reporter with the supernatant solution induced the AmiE expression was almost the same as the one with the supernatant solution not induced the AmiE expression.(Fig.1)

However, in case of 3OC12HSL, the fluorescence intensity of the reporter added the solution induced the AmiE expression was about 1/10 of the reporter added the solution not induced AmiE expression. Based on the above, it is concluded that AmiE does not degrade C4HSL but degrades 3OC12HSL.(Fig.2)



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961