Difference between revisions of "Part:BBa K1897018:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | There | + | There are HA tag(s) available for characterisation of the LuxR, LuxI and invasin proteins respectively produced via western blot. Note that the stop codons for LuxR <partinfo>BBa_C0062</partinfo>, LuxI <partinfo>BBa_C0061</partinfo> and Invasin <partinfo>BBa_K1897010</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=== | ===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>. | ||
− | + | 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>. | |
+ | |||
+ | Invasin is derived from <i>Yersinia pseudotuberculosis</i>, from the coding sequence [https://www.ncbi.nlm.nih.gov/nuccore/M17448.1 [Genbank M17448.1]] (see <partinfo>BBa_K1897010</partinfo>. 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=== | ===References=== | ||
+ | hoi, S. H., & Greenberg, E. P. (1992). Genetic evidence for multimerization of LuxR, the transcriptional activator of Vibrio fischeri luminescence. <i>Molecular marine biology and biotechnology, 1</i>(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. <i>Journal of bacteriology, 176</i>(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. <i>Journal of bacteriology, 173</i>(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). |
Revision as of 20:51, 15 October 2016
LuxR + LuxI + Invasin
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1852
Illegal BamHI site found at 1872
Illegal XhoI site found at 4659 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 50
Illegal AgeI site found at 1065
Illegal AgeI site found at 2844
Illegal AgeI site found at 4353 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
There are HA tag(s) available for characterisation of the LuxR, LuxI and invasin proteins respectively produced via western blot. Note that the stop codons for LuxR BBa_C0062, LuxI BBa_C0061 and Invasin BBa_K1897010 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.
Invasin is derived from Yersinia pseudotuberculosis, from the coding sequence [Genbank M17448.1] (see BBa_K1897010. 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
hoi, 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).