Difference between revisions of "Part:BBa K1632004"
Line 4: Line 4:
 
[[Image:Tokyo Tech fim switch (wild-type) design.png|thmub|center|400px|Fig. 1. Overview of fim inversion system]]<br>
 
[[Image:Tokyo Tech fim switch (wild-type) design.png|thmub|center|400px|Fig. 1. Overview of fim inversion system]]<br>
  
−
''Fim'' switch (wild-type) is derived from wild-type sequence. ''Fim'' switch (wild type) have sigma 70 promoter which is constitutively promoter. The promoter in the ''fim'' switch transcripts to the right. From the direction of transcription, the state is defined as ON. On the other hand, fim switch [Default ON](wild-type)(<partinfo>BBa_K1632005</partinfo>) transcripts to the right.<br>
+
''Fim'' switch (wild-type) is derived from wild-type sequence. ''Fim'' switch (wild-type) have sigma 70 promoter which is constitutively promoter. The promoter in the ''fim'' switch transcripts to the right. From the direction of transcription, the state is defined as ON. On the other hand, fim switch [Default ON](wild-type)(<partinfo>BBa_K1632005</partinfo>) transcripts to the right.<br>
  
  

Revision as of 19:26, 13 September 2015

fim switch[default ON](wild-type)

Fig. 1. Overview of fim inversion system

Fim switch (wild-type) is derived from wild-type sequence. Fim switch (wild-type) have sigma 70 promoter which is constitutively promoter. The promoter in the fim switch transcripts to the right. From the direction of transcription, the state is defined as ON. On the other hand, fim switch [Default ON](wild-type)(BBa_K1632005) transcripts to the right.


Fim switch is inverted by two recombinase, FimB and FimE.The FimB protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction with approximately equal efficiencies. On the other hand, the FimE protein inverts fim switch predominantly in the ON-to-OFF direction.


We constructed fim switch[ON](wild-type)-rbs-gfp(BBa_K1632007) to characterize the function of this part, by inserting this promoter upstream of a GFP coding sequence.

In the fimB dependent fim switch state assay, we transformed fim switch[ON](wild-type)-rbs-gfp and pBAD/araC-fimB(BBa_K1632012)in the E.coli DH5alfa strain. We measured the fluorescence intensity of the cells induced by arabinose.

Similary,in the fimE dependent fim switch state assay, we transformed fim switch[ON](wild-type)-rbs-gfp and pBAD/araC-fimE(BBa_K1632013) in the E.coli DH5alfa strain.We measured the fluorescence intensity of the cells induced by arabinose.


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]