Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Graeme Glaister
Date created: 2015-09-16 11:00:00
Date modified: 2015-09-23 01:34:01
Low RBS expressing MS2 Coat protein ribozyme affinity purification (RAP)
| Types | DnaRegion |
| Roles | DNA sequence_feature |
| Sequences | BBa_K1791002_sequence (Version 1) |
Description
low RBS expressing MS2 Coat protein ribozyme affinity purification (RAP). This part utilizes the expression of C-terminal 6X His tagged fusion MS2 coat protein under a T7 promoter and a low efficiency shine dalgarno sequence for expression of MS2. Downstream there is a T7 promoter followed by a theophylline ribozyme and a MS2 apatamer at the 3' end. Co-expression of the two modules results in MS2 binding to the 3' end of the theophylline ribozyme transcript allowing for the MS2-RNA purification by nickel affinity chromatography. The RNA can be eluted from the complex by the addition of theophylline.Notes
We had to consider MS2 expression levels relative to the theophylline ribozyme transcript and assessed how this affects total RNA yield. In order to test this we developed a second part with increased MS2 expression (BBa_K1791001)Source
This is an amalgamation of several biobricks an MS2 coat protein, an MS2 binding domain and the theophylline ribozyme (BBa_K1791000)| Access | Instance | Definition |
| public public public public public public public public | BBa_I719005 BBa_B0031 BBa_K1791003 BBa_B0015 BBa_K1791007 BBa_K1791005 BBa_K1791008 BBa_K1791006 | pT7 BBa_B0031 BBa_K1791003 BBa_B0015 BBa_K1791007 BBa_K1791005 BBa_K1791008 BBa_K1791006 |