Difference between revisions of "Part:BBa J100066"
(New page: This part is a synthetic riboswitch that can be used in E. coli. In the presence of theophylline, the expression of the gene of interest is induced, but in the absence of theophylline, ver...) |
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Because the riboswitch must be right beside the gene of interest (the start codon is folded in the riboswitch), we added a BsaI site. This allows the riboswitch to be attached to the gene of interest using Golden Gate Assembly (as long as the first nucleotide after the start codon of the gene of interest is a C, we designed the part to use with superfolder GFP). | Because the riboswitch must be right beside the gene of interest (the start codon is folded in the riboswitch), we added a BsaI site. This allows the riboswitch to be attached to the gene of interest using Golden Gate Assembly (as long as the first nucleotide after the start codon of the gene of interest is a C, we designed the part to use with superfolder GFP). | ||
| − | This riboswitch is modified from riboswitch E as described in the paper by Topp et al. | + | This riboswitch is modified from riboswitch E as described in the paper by [[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2988590/?tool=pubmed Topp ''et al''.]] |
| − | Compare this part with [https://parts.igem.org/Part:BBa_J100065 Riboswitch D] | + | Compare this part with [https://parts.igem.org/Part:BBa_J100065 Riboswitch D J100065]. Riboswitch E has a longer RBS. See Figure 1.B in Topp "et al" for the comparison of gene expression. |
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| + | This part has been combined with superfolder GFP using the BsaI sites ([https://parts.igem.org/Part:BBa_J100080 Part J100080). | ||
References: Shana Topp, Colleen M. K. Reynoso, Jessica C. Seeliger, Ian S. Goldlust, Shawn K. Desai, Dorothée Murat, Aimee Shen, Aaron W. Puri, Arash Komeili, Carolyn R. Bertozzi, June R. Scott, and Justin P. Gallivan. Synthetic Riboswitches That Induce Gene Expression in Diverse Bacterial Species. Applied and Environmental Microbiology, 76:23, 7881-7884. December 2010. | References: Shana Topp, Colleen M. K. Reynoso, Jessica C. Seeliger, Ian S. Goldlust, Shawn K. Desai, Dorothée Murat, Aimee Shen, Aaron W. Puri, Arash Komeili, Carolyn R. Bertozzi, June R. Scott, and Justin P. Gallivan. Synthetic Riboswitches That Induce Gene Expression in Diverse Bacterial Species. Applied and Environmental Microbiology, 76:23, 7881-7884. December 2010. | ||
Latest revision as of 14:26, 12 July 2012
This part is a synthetic riboswitch that can be used in E. coli. In the presence of theophylline, the expression of the gene of interest is induced, but in the absence of theophylline, very little transcription takes place. This part contains a modified T5 promoter, which, in the absence of cymR, acts as a strong constitutive promoter. The ribosomal binding site is contained in the riboswitch. Because the riboswitch must be right beside the gene of interest (the start codon is folded in the riboswitch), we added a BsaI site. This allows the riboswitch to be attached to the gene of interest using Golden Gate Assembly (as long as the first nucleotide after the start codon of the gene of interest is a C, we designed the part to use with superfolder GFP).
This riboswitch is modified from riboswitch E as described in the paper by http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2988590/?tool=pubmed Topp ''et al''.
Compare this part with Riboswitch D J100065. Riboswitch E has a longer RBS. See Figure 1.B in Topp "et al" for the comparison of gene expression.
This part has been combined with superfolder GFP using the BsaI sites ([https://parts.igem.org/Part:BBa_J100080 Part J100080).
References: Shana Topp, Colleen M. K. Reynoso, Jessica C. Seeliger, Ian S. Goldlust, Shawn K. Desai, Dorothée Murat, Aimee Shen, Aaron W. Puri, Arash Komeili, Carolyn R. Bertozzi, June R. Scott, and Justin P. Gallivan. Synthetic Riboswitches That Induce Gene Expression in Diverse Bacterial Species. Applied and Environmental Microbiology, 76:23, 7881-7884. December 2010.