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]

Shipped as purified and dried down plasmid DNA stained with Cresol Red in one 96-well plate. Each well contains approximately 50 ng of DNA. Cresol Red will not impact plasmid transformation.

A plate map can be found here.

Where can I find more information –

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 – 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 – 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
mehmet tardu	mtardu {at} gmail {dot} com	Turkey
Zoila E Jurado Quiroga	zjuradoq {at} caltech {dot} edu	United States
Jennifer Molloy	jcm80 {at} cam {dot} ac {dot} uk	United Kingdom
Nicholas White	nickk.white {at} gmail {dot} com	United States
Jordan Gonzalez	jgonzalez {at} thecitizensciencelab {dot} org	United States
Benjamin Arias	barias {at} alumni {dot} usfq {dot} edu {dot} ec	Ecuador
David Peabody	dpeabody {at} salud {dot} unm {dot} edu	United States
Piero Beraun	piero.beraun {at} utec {dot} edu {dot} pe	Italy
Shobha KrupaRani	pshobha {at} ccmb {dot} res {dot} in	India
Aleksandr Shilovich	mercurialbadger {at} yandex {dot} ru	Russia
Rodolfo Marcolino	rodolfo.marcolino {at} me {dot} com	Argentina
Ian Cubit	Djcubit {at} nycap {dot} rr {dot} com	United States
Juan Sebastian Alvarez	juan.alvarez1 {at} ucalgary {dot} ca	Canada
Amjed Alsultan	NA	Iraq
Zoila E Jurado Quiroga	mikiy {at} caltech {dot} edu	United States
Yan Zhang	yz473 {at} gatech {dot} edu	United States

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

Back to Endy Lab at Stanford University

MTA

Unilateral OpenMTA

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