PUREiodic Table Construction Kit

PUREiodic Table Construction Kit

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What is it

The construction of synthetic cells from lifeless ensembles of molecules is expected to require integration of hundreds of genetically-encoded functions whose collective capacities enable self-reproduction in simple environments. Here, we provide a partial toolkit for systematically evaluating the capacity of a system to remake itself. Using the cell-free Protein synthesis Using Recombinant Elements (PURE) as a model system one may evaluate the capacity of PURE, whose composition is completely known, to remake 36 life-essential functions. 

The PUREiodic Table Construction Kit contains 36 single-components of the PURE cell-free system in pET28a vector backbones. Each component is expressible in E. coli with T7 RNA polymerase and can be His purified. 


How to Use the PUREiodic Table Construction Kit

Researchers can use the kit to systematically determine and quantify which individual enzymatic components of PURE, when removed from PURE, result in diminished PURE activity as measured by expression of a reporter gene. Since the standard commercially-available starting material (PURExpress) combines all of the enzymes into a single tube (PURExpress Solution B), one may produce their own custom mixtures consisting of single-enzyme depletions by purifying each enzyme comprising PURE and reconstituting single-enzyme depletion PURE variants. 

Researchers can also perform complementation experiments by supplementing each single-component depletion with a cognate PURE-made component and observing if and to what extent PURE activity is recovered. 

We represent our previous work with the kit via a standard visual form we call the PUREiodic Table that serves as a tool for tracking which life-essential functions can work together in remaking one another and what functions remain to be remade.[1]

All parts are present on the plate as raw DNA.

A plate map can be found here.


Where can I find more information –

  1. https://doi.org/10.1101/2021.03.03.433818

Genes

Gene Name Freegenes ID
Inorganic Pyrophosphotase N-tagged Inorganic Pyrophosphotase N-tagged BBF10K_000545
Methionine-tRNA ligase C-tagged Methionine-tRNA ligase C-tagged BBF10K_000547
pET-28a – AlaRS pET-28a_(+)_AlaRS BBF10K_000549
pET-28a – ArgRS pET-28a_(+)_ArgRS BBF10K_000550
pET-28a – AsnRS pET-28a_(+)_AsnRS BBF10K_000551
pET-28a – AspRS pET-28a_(+)_AspRS BBF10K_000552
pET-28a – CK pET-28a_(+)_CK BBF10K_000553
pET-28a – CysRS pET-28a_(+)_CysRS BBF10K_000554
pET-28a – EF-G pET-28a_(+)_EF-G BBF10K_000555
pET-28a – EF-Ts pET-28a_(+)_EF-Ts BBF10K_000556
pET-28a – EF-Tu pET-28a_(+)_EF-Tu BBF10K_000557
pET-28a – GlnRS pET-28a_(+)_GlnRS BBF10K_000558
pET-28a – GluRS pET-28a_(+)_GluRS BBF10K_000559
pET-28a – GlyRS pET-28a_(+)_GlyRS BBF10K_000560
pET-28a – HisRS pET-28a_(+)_HisRS BBF10K_000561
pET-28a – IF1 pET-28a_(+)_IF1 BBF10K_000562
pET-28a – IF2 pET-28a_(+)_IF2 BBF10K_000563
pET-28a – IF3 pET-28a_(+)_IF3 BBF10K_000564
pET-28a – IleRS pET-28a_(+)_IleRS BBF10K_000565
pET-28a – LeuRS pET-28a_(+)_LeuRS BBF10K_000567
pET-28a – LysRS pET-28a_(+)_LysRS BBF10K_000568
pET-28a – MK pET-28a_(+)_MK BBF10K_000570
pET-28a – MTF pET-28a_(+)_MTF BBF10K_000575
pET-28a – NDK pET-28a_(+)_NDK BBF10K_000576
pET-28a – PheRS pET-28a_(+)_PheRS BBF10K_000577
pET-28a – ProRS pET-28a_(+)_ProRS BBF10K_000578
pET-28a – RF1 pET-28a_(+)_RF1 BBF10K_000579
pET-28a – RF3 pET-28a_(+)_RF3 BBF10K_000581
pET-28a – RRF pET-28a_(+)_RRF BBF10K_000582
pET-28a – SerRS pET-28a_(+)_SerRS BBF10K_000583
pET-28a – T7RNAP pET-28a_(+)_T7RNAP BBF10K_000584
pET-28a – ThrRS pET-28a_(+)_ThrRS BBF10K_000585
pET-28a – TrpRS pET-28a_(+)_TrpRS BBF10K_000586
pET-28a – TyrRS pET-28a_(+)_TyrRS BBF10K_000587
pET-28a – ValRS pET-28a_(+)_ValRS BBF10K_000588

Download all of this information as a CSV from our GitHub.

Bionet

The bionet enables open peer-peer exchange of functional biomaterials and associated data. This product may also be available from bionet nodes that are more convenient to you. Here are other bionet nodes who may be willing to provide you this specific product.

Name Contact Country
Rodolfo Marcolino rodolfo.marcolino {at} me {dot} com Argentina
Zoila E Jurado Quiroga zjuradoq {at} caltech {dot} edu United States
Jordan Gonzalez jgonzalez {at} thecitizensciencelab {dot} org United States

Download all of this information as a CSV from our GitHub.
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