	US 11,866,712 B2
	System based on the reassembly of GFP for studying the trans-translational activity and identifying new antibiotics
Reynald Gillet, Le Verger (FR); and Charlotte Guyomar, Porspoder (FR)
Assigned to UNIVERSITE DE RENNES 1, Rennes (FR); and CENTRE NATIONALE DE LA RECHERCHE SCIENTIFIQUE—CNRS, Paris (FR)
Appl. No. 17/056,574
Filed by UNIVERSITE DE RENNES 1, Rennes (FR); and CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE—CNRS—, Paris (FR)
PCT Filed May 25, 2018, PCT No. PCT/EP2018/063780 § 371(c)(1), (2) Date Nov. 18, 2020, PCT Pub. No. WO2019/223879, PCT Pub. Date Nov. 28, 2019.
Prior Publication US 2021/0207151 A1, Jul. 8, 2021
Int. Cl. C12N 15/70 (2006.01); G01N 33/58 (2006.01); C12N 15/74 (2006.01); C12N 15/78 (2006.01); C12Q 1/02 (2006.01); C12Q 1/18 (2006.01); G01N 21/64 (2006.01)

CPC C12N 15/70 (2013.01) [C12N 15/74 (2013.01); C12N 15/746 (2013.01); C12N 15/78 (2013.01); C12Q 1/025 (2013.01); C12Q 1/18 (2013.01); G01N 21/6486 (2013.01)]

28 Claims

1. A reporter system for the trans-translational activity of a bacterial species selected from the group consisting of Escherichia coli, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter cloacae, and Mycobacterium tuberculosis, comprising:

a nucleotide sequence encoding the first 10 domains of GFP and not comprising a stop codon;

a nucleotide sequence encoding the SmpB protein of said bacterial species; and

a modified tmRNA, wherein the modified tmRNA is the tmRNA of said bacterial species, in which the portion of sequence encoding the proteolysis tag is replaced with a nucleotide sequence consisting of sequence SEQ ID NO: 19 which encodes the 11^thdomain of GFP of sequence SEQ NO: 18.