Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Abiel Trevi??o Garza
Date created: 2012-09-23 11:00:00
Date modified: 2015-05-08 01:13:34
pBAD-pXyl AND Gate
| Types | DnaRegion |
| Roles | Regulatory promoter |
| Sequences | BBa_K851002_sequence (Version 1) |
Description
pBAD-pXyl is a combined promoter of D-Xylose and L-arabinose sugar sensor systems which is designed to be activated only in the presence of both sugars in the medium therefore functioning as an AND logic gate.BIOLOGY
pXyl is an inducible promoter regulated by the transcriptional regulator XylR which, in Bacillus subtilis, regulates the expression of xyl operon[1]. Gene expression under pXyl can be induced by the addition of D-Xylose to the medium [1, 2]. The nucleotide sequence was obtained from Wilhelm, M &C. P. Hollenberg, 1985[3].
In the presence of L-arabinose, expression from pBAD is turned on while the absence of L-arabinose produces very low levels of transcription from pBAD [4, 5]. More precisely, in the absence of arabinose, the repressor protein AraC (BBa_I13458[6]) binds to the AraI1 operator site of pBAD and the upstream operator site AraO2, blocking transcription[7], but in the presence of arabinose, AraC binds to it and changes its conformation such that it interacts with the AraI1 and AraI2 operator sites, permitting transcription[7]. The nucleotide sequence was similar of that in Part:BBa_K206000[8].
For iGEM UNAM Genomics M??xico 2012 project [9], pBAD/pXyl was used in the design of an AND logic gate[10] using a recently described new type of communication system between Bacillus Subtilis cells called Nanotubes[11].
Notes
chimeric promotorSource
REFERENCES[1] D Gartner, M Geissendorfer, & W Hillen(1988). Expression of the Bacillus subtilis xyl Operon Is Repressed at the Level of Transcription and Is Induced by Xylose J Bacteriol 170:7,3102-3109.
[2] Shamanna, D. K., and K. E. Sanderson. 1979. Genetics and regulation of D-xylose utilization in Salmonella typhimurium LT2. J. Bacteriol. 139:71-79.
[3] Wilhelm, M., and C. P. Hollenberg. 1985. Nucleotide sequence of the Bacillus subtilis xylose isomerase gene: extensive homology between the Bacillus and E. coli enzyme. Nucleic Acids Res. 13:5717-5722.
[4] Lee, N. (1980) Molecular Aspects of ara Regulation. In The Operon, J. H. Miller and W. S. Reznikoff, eds. (Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory), pp. 389-410.
[5] Lee, N., Francklyn, C., and Hamilton, E. P. (1987). Arabinose-Induced Binding of AraC Protein to araI2 Activates the araBAD Operon Promoter. Proc. Natl. Acad. Sci. USA 84, 8814-8818.
[6] http://partsregistry.org/wiki/index.php/Part:BBa_I13458
[8] http://partsregistry.org/Part:BBa_K206000
[7] Schlief, R. (2000). Regulation of the L-arabinose operon of Escherichia coli. Trends in Genetics. 16(12):559-565.
[9] http://2012.igem.org/Team:UNAM_Genomics_Mexico
[10] http://2012.igem.org/Team:UNAM_Genomics_Mexico/Project/Description
[11] Dubey GP, Ben-Yehuda S. (2011) Intercellular nanotubes mediate bacterial communication. Cell.;144(4):590-600
[12] Guzman, L.-M., Belin, D., Carson, M. J., and Beckwith, J. (1995). Tight Regulation, Modulation, and High-Level Expression by Vectors Containing the Arabinose PBAD Promoter. J. Bacteriol. 177, 4121-4130.
[13] David, J. D., and H. Weismeyer. 1970. Control of xylose metabolism in Escherichia coli. Biochim. Biophys. Acta 201: 497-499.
| Sequence Annotation | Location | Component / Role(s) |
| pBad recognition region 1 Pbad recognition region 2 -35 -10 XylR Binding Sites XylR Binding Site 2 | 3,19 161,174 247,253 271,276 288,299 305,313 | feature/binding non_covalent_binding_site non_covalent_binding_site feature/binding feature/binding non_covalent_binding_site feature/binding non_covalent_binding_site non_covalent_binding_site feature/binding non_covalent_binding_site feature/binding |