Difference between revisions of "Part:BBa K1613013"

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It can produce Laccase which can degrade benzo[a]pryene and QsrR which can combine with bap-1,6-quinone.
 
It can produce Laccase which can degrade benzo[a]pryene and QsrR which can combine with bap-1,6-quinone.
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 +
<p style="font-size:150%">'''Gel Electrophoresis'''</p>
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[[File:HSNU-TAIPEI-BBa_K1613013.jpg]]
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Fig.1:The Gel Electrophoresis picture of BBa_K1613013. The plasmid was cut by restriction enzymes EcoRI and SpeI. There are three bands in the picture: Plasmids, Backbones, and Parts.
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<p style="font-size:150%">'''E.Coli Survival'''</p>
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To ensure the biosensor can work in the toxin solution. We did experiment of E.coli survival. To make sure the toxin won't kill it.
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 +
<p style="font-size:120%">'''Procedure'''</p>
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First we culture DH5α with LB only plate for 15hr. Then,pick one colony in the LB broth,and liquid culture for 15hr.
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Take 80μL into 2ml LB broth × 6 tubes and then culture 1 hr.
 +
After 1hr,add 20μL benzo[a]pryene into three tubes(conc. Is 2000ppb(A thousand times the standard value))
 +
And add 20μL DMSO into the other tubes.Then,culture for 3hr.
 +
After 3hr,dilute the broth to 10-6
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And take 200μL to spread the plate.
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 +
▼Table1: E. coli on the agar plate.
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 +
[[File:HSNU-TAIPEI-BZP-904-1.jpg]]
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<p style="font-size:120%">'''Results'''</p>
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▼Table2: E. coli on the agar plate.
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 +
[[File:HSNU-TAIPEI-BZP-820-4.jpg]]
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 +
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[[File:HSNU-TAIPEI-BZP-904-2.jpg]]
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▲Fig2: Benzo[a]pyrene Category A
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Benzo[a]pryene Category A
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[[File:HSNU-TAIPEI-BZP-904-3.jpg]]
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▲Fig3:Control Category A
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<p style="font-size:150%">'''E.coli in the Microcapsules'''</p>
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We designed a microcapsule device to keep E.coli in it.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 03:35, 26 September 2015

Laccase and Quinone-sensing and response Repressor(QsrR)

It can produce Laccase which can degrade benzo[a]pryene and QsrR which can combine with bap-1,6-quinone.

Gel Electrophoresis

HSNU-TAIPEI-BBa K1613013.jpg

Fig.1:The Gel Electrophoresis picture of BBa_K1613013. The plasmid was cut by restriction enzymes EcoRI and SpeI. There are three bands in the picture: Plasmids, Backbones, and Parts.

E.Coli Survival

To ensure the biosensor can work in the toxin solution. We did experiment of E.coli survival. To make sure the toxin won't kill it.

Procedure

First we culture DH5α with LB only plate for 15hr. Then,pick one colony in the LB broth,and liquid culture for 15hr. Take 80μL into 2ml LB broth × 6 tubes and then culture 1 hr. After 1hr,add 20μL benzo[a]pryene into three tubes(conc. Is 2000ppb(A thousand times the standard value)) And add 20μL DMSO into the other tubes.Then,culture for 3hr. After 3hr,dilute the broth to 10-6 And take 200μL to spread the plate.

▼Table1: E. coli on the agar plate.

HSNU-TAIPEI-BZP-904-1.jpg

Results

▼Table2: E. coli on the agar plate.

HSNU-TAIPEI-BZP-820-4.jpg


HSNU-TAIPEI-BZP-904-2.jpg

▲Fig2: Benzo[a]pyrene Category A

Benzo[a]pryene Category A

HSNU-TAIPEI-BZP-904-3.jpg

▲Fig3:Control Category A

E.coli in the Microcapsules

We designed a microcapsule device to keep E.coli in it.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1409
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1570