Difference between revisions of "Part:BBa K1949050"

 
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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 six carbons.
 
<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 six carbons.
  
 
[[Image:amiE1.png|thumb|center|400px| Fig.1  AmiE is an acylase that degrades long chain N-acyl homoserine lactone (AHL) molecules with acyl chains longer than eightsix carbons]]<br>
 
  
 
===Characterization===
 
===Characterization===
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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.
  
・PBAD/araC-<i>rbs-amiE</i>(pSB6A1)
+
・PBAD/araC - <i>rbs - amiE</i>(pSB6A1)
  
・Ptet-<i>rbs-lux</i>R-<i>TT</i>-Plux-<i>rbs-gfp</i> (pSB6A1)
+
・Ptet - <i>rbs - lux</i>R - <i>tt</i> - Plux - <i>rbs - gfp</i> (pSB6A1)
  
・Ptet-<i>rbs-rhl</i>R-<i>TT</i>-Plux-<i>rbs-gfp</i> (pSB6A1)
+
・Ptet - <i>rbs - rhl</i>R - <i>tt</i> - Plux - <i>rbs - gfp</i> (pSB6A1)
  
[[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>
+
[[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>
  
[[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>
+
[[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.

Latest revision as of 15:07, 31 October 2017


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 six 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
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]