Part:BBa_K1613013

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

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.

Results

Table1: E. coli on the agar plate.

Influence of bento[a]pyrene on E.coli(DH5α) survival rate
Group	Colony
Benzo[A]Pyrene	237	201	104
Control	189	51	233

Fig2: Benzo[a]pyrene Group

Fig3:Control Group

There is no difference in amount of E.coli between two groups which means benzopyrene will not affect the survival of E.coli.

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