Voigt Lab Fragmented T7 RNAP Plasmids
Christopher Voigt's lab has built a ‘resource allocator’ to control synthetic genetic systems by splitting T7 RNA polymerase (RNAP) variants into a number of fragments. By tuning the production of a limiting protein fragment, the total number of polymerases in a synthetic system can be controlled while allowing the allocation of this resource between different outputs.
A publication from the Voigt lab in Molecular Systems Biology (Segall-Shapiro TH, et al., MSB, 2014) describes the bisection mapping of T7 RNAP to produce functional protein fragments and the composition of these fragments into a resource allocator system. Four orthogonal T7 RNAP variants were bisected into a conserved ‘core fragment’ and variable ‘σ fragments’. T7 RNAP is further shown to tolerate fragmentation into three and four fragments while retaining function. The three-piece RNAP and further mutations are used to create accessory fragments for further regulation of the resource allocator.
The work and corresponding plasmids are described in:
The Voigt lab has deposited plasmids encoding the fragmented T7 RNAP system that can be used to implement resource allocation. These include orthogonal T7 RNAP variant reporter plasmids (pTHSSd_8 – 11), plasmids with inducible σ fragments (pTHSSd_23 – 26), core fragment expression plasmids (pTHSSd_38 – 40), ‘accessory’ fragments (pTHSSd_27, 29, 42), and control plasmids (pTHSSd_34, 41, 44). They have also deposited plasmids to express three- and four-piece T7 RNAP (pTHSSd_14, 18) and a modified transposon that can be used to map the tolerated bisection sites in a protein (pTHSSd_1).
Individual plasmids can be ordered via the links below:
|59947||pTHSSd_1||Add to Cart|
|59948||pTHSSd_8||Add to Cart|
|59949||pTHSSd_9||Add to Cart|
|59951||pTHSSd_11||Add to Cart|
|59952||pTHSSd_14||Add to Cart|
|59953||pTHSSd_18||Add to Cart|
|59954||pTHSSd_23||Add to Cart|
|59955||pTHSSd_24||Add to Cart|
|59956||pTHSSd_25||Add to Cart|
|59957||pTHSSd_26||Add to Cart|
|59958||pTHSSd_27||Add to Cart|
|59959||pTHSSd_29||Add to Cart|
|59960||pTHSSd_34||Add to Cart|
|59963||pTHSSd_40||Add to Cart|
|59966||pTHSSd_44||Add to Cart|