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.