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Revision as of 05:36, 19 October 2017

Composite Parts

Part BBa_K1791001

Part BBa_K1791001 contains two modules, a standard ribosome binding site (RBS) expressing MS2 coat protein and a theophylline ribozyme with a MS2 aptamer at the 3’ end. To express MS2 coat protein the part utilizes expression of c-terminal 6x His tagged MS2 coat protein under the regulation of a T7 promoter and a standard Shine Dalgarno sequence followed by a double terminator (BBa_0015). Downstream of the RBS is a T7 promoter followed by a theophylline ribozyme and a MS2 aptamer at the 3’ end. Coexpression of the two modules results in MS2 binding to the 3’ end of the theophylline ribozyme module which allows for MS2-RNA purification by nickel affinity chromatography, RNA can then be eluted from the MS2-RNA complex by addition of theophylline.

<img src="Uleth15_StandardRPA.png" height="30%" width="30%">

Check it out on the <a href="http://parts.igem.org/Part:BBa_K1791001">Registry</a>

Part BBa_K1791002

Part BBa_K17910012 contains two modules, a low efficiency ribosome binding site (RBS) expressing MS2 coat protein and a theophylline ribozyme with a MS2 aptamer at the 3’ end. To express MS2 coat protein the part utilizes expression of c-terminal 6x His tagged MS2 coat protein under a T7 promoter and a standard Shine Dalgarno sequence followed by a double terminator (BBa_0015). Downstream of the RBS is a T7 promoter followed by a theophylline ribozyme and a MS2 aptamer at the 3’ end. Coexpression of the two modules results in MS2 binding to the 3’ end of the theophylline ribozyme construct which allows for MS2-RNA purification by nickel affinity chromatography, RNA can then be eluted from the MS2-RNA complex by addition of theophylline.

<img src="Uleth_ConstructSchematic.png" height="30%" width="30%">

Check it out on the <a href="http://parts.igem.org/Part:BBa_K1791002">Registry</a>

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