PXD066302

PXD066302 is an original dataset announced via ProteomeXchange.

Dataset Summary

Title	Nanoluc synthesized by the PURE system with the tRNA array method
Description	Transfer RNA (tRNA) plays a central role in translation. Simultaneously synthesizing the minimal yet sufficient number of tRNA species (at least 21) in vitro poses a challenge to constructing a self-reproducible artificial cell. In this study, we developed a simplified processing method that enables the simultaneous expression of all 21 tRNAs in a reconstituted transcription/translation system (PURE system). By combining direct tRNA linkage and HDVR attachment methods, termed the tRNA array method, we achieved simultaneous expression of all 21 tRNAs from a single polycistronic DNA template in the PURE system. This method allows for the easy customization of the genetic code by introducing additional tRNAs. To demonstrate genetic code reassignment, we replaced five Leu residues (CUU) in Nanoluc with ACG, which is assigned to Thr in the native codon table. To confirm amino acid reassignment, we performed LC-MS/MS analysis of translated Nanoluc proteins. The results confirmed the incorporation of threonine at the reassigned ACG codon in the presence of tRNA^Thr_CGU, and leucine incorporation when using tRNA^Leu_CGU.
HostingRepository	jPOST
AnnounceDate	2025-07-25
AnnouncementXML	Submission_2025-07-24_23:13:51.553.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Keiko Masuda
SpeciesList	scientific name: Acanthephyra purpurea; NCBI TaxID: 576170;
ModificationList	S-carboxamidomethyl-L-cysteine; L-methionine sulfoxide
Instrument	LTQ Orbitrap Velos

Dataset History

Revision	Datetime	Status	ChangeLog Entry
0	2025-07-18 02:11:22	ID requested
⏵ 1	2025-07-24 23:13:51	announced

Publication List

Dataset with its publication pending

Dataset with its publication pending

Keyword List

submitter keyword: PURE system, tRNA array, genetic code-engineering

submitter keyword: PURE system, tRNA array, genetic code-engineering

Contact List

Norikazu Ichihashi
lab head
Keiko Masuda
contact affiliation	RIKEN BDR
dataset submitter

Full Dataset Link List

jPOST dataset URI
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jPOST dataset URI

Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.jpostdb.org/JPST003947/