Difference between revisions of "Part:BBa K1897015:Design"

Latest revision as of 17:59, 15 October 2016

LuxR + LuxI

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
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1852
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 50
Illegal AgeI site found at 1065
1000
COMPATIBLE WITH RFC[1000]

Design Notes

There is one HA tag available for characterisation of the LuxR and LuxI proteins respectively produced via western blot. Note that the stop codons for LuxR BBa_C0062 and LuxI BBa_C0061 is shifted to after the HA tag.

Also, the transcriptional terminators rrnBT1 (from BBa_B0010) + BBa_B0012 for LuxR was derived from BBa_B0015, with the first 8 base pairs removed.

Source

LuxR was obtained from biobrick part BBa_C0062, which was derived from Vibrio fischeri. The promoter was synthesized based on the sequence obtained from BBa_J23119. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

LuxI was obtained from biobrick part BBa_C0061, which was derived from Vibrio fischeri. The promoter was synthesized based on the sequence obtained from BBa_R0062. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

References

Choi, S. H., & Greenberg, E. P. (1992). Genetic evidence for multimerization of LuxR, the transcriptional activator of Vibrio fischeri luminescence. Molecular marine biology and biotechnology, 1(6), 408-413.

Fuqua, W. C., Winans, S. C., & Greenberg, E. P. (1994). Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators. Journal of bacteriology, 176(2), 269.

Shadel, G. S., & Baldwin, T. O. (1991). The Vibrio fischeri LuxR protein is capable of bidirectional stimulation of transcription and both positive and negative regulation of the luxR gene. Journal of bacteriology, 173(2), 568-574.

Trott, A. E. (2000). Amino Acid Residues in LuxR Critical for its Mechanism of Transcriptional Activation during Quorum Sensing (Doctoral dissertation, Virginia Polytechnic Institute and State University).

@@ Line 7: / Line 7: @@
 ===Design Notes===
-There is one HA tag available for characterisation of the LuxR and LuxI proteins respectively produced via western blot.
+There is one HA tag available for characterisation of the LuxR and LuxI proteins respectively produced via western blot. Note that the stop codons for LuxR <partinfo>BBa_C0062</partinfo> and LuxI <partinfo>BBa_C0061</partinfo> is shifted to after the HA tag.
+Also, the transcriptional terminators rrnBT1 (from <partinfo>BBa_B0010</partinfo>) + <partinfo>BBa_B0012</partinfo> for LuxR was derived from <partinfo>BBa_B0015</partinfo>, with the first 8 base pairs removed.
 ===Source===
+LuxR was obtained from biobrick part <partinfo>BBa_C0062</partinfo>, which was derived from <i>Vibrio fischeri</i>.
+The promoter was synthesized based on the sequence obtained from <partinfo>BBa_J23119</partinfo>.
+The ribosome binding site (RBS) was synthesized based on the sequence obtained from <partinfo>BBa_B0030</partinfo>.
+The transcription terminators were synthesized based on the sequence obtained from <partinfo>BBa_B0015</partinfo>.
-These parts are derived from <i>Vibrio fischeri</i>.
+LuxI was obtained from biobrick part <partinfo>BBa_C0061</partinfo>, which was derived from <i>Vibrio fischeri</i>. The promoter was synthesized based on the sequence obtained from <partinfo>BBa_R0062</partinfo>. The ribosome binding site (RBS) was synthesized based on the sequence obtained from <partinfo>BBa_B0030</partinfo>. The transcription terminators were synthesized based on the sequence obtained from <partinfo>BBa_B0015</partinfo>.
 ===References===