A GRA2 minimal promoter improves the efficiency of TATi / Tet-Off conditional regulation of gene expression in Toxoplasma gondii.
Antigens, Protozoan
/ genetics
Bacterial Proteins
/ genetics
Cells, Cultured
Cloning, Molecular
/ methods
Fibroblasts
/ parasitology
Gene Expression
Gene Expression Regulation
/ drug effects
Genes, Reporter
Humans
Luminescent Proteins
/ genetics
Membrane Glycoproteins
/ genetics
Plasmids
/ chemistry
Promoter Regions, Genetic
Protozoan Proteins
/ genetics
Recombinant Proteins
/ chemistry
Tetracyclines
/ pharmacology
Toxoplasma
/ drug effects
Transcription, Genetic
Transfection
Apicomplexa
Gene control
Improvement
Minimal promoter
Regulation
Signal to Noise Ratio
TATi
Tet-Off
Transcription
Journal
Molecular and biochemical parasitology
ISSN: 1872-9428
Titre abrégé: Mol Biochem Parasitol
Pays: Netherlands
ID NLM: 8006324
Informations de publication
Date de publication:
07 2021
07 2021
Historique:
received:
22
03
2021
revised:
23
05
2021
accepted:
24
05
2021
pubmed:
30
5
2021
medline:
2
9
2021
entrez:
29
5
2021
Statut:
ppublish
Résumé
A tetracycline-responsive transcription system (Tet-Off) adapted for use in Toxoplasma gondii (nicknamed TATi) is useful for molecular biological studies of this organism. Previous studies using TATi incorporated minimal promoters derived from the gene promoters for TgSAG1 or TgSAG4. The present study achieves improved activation and suppression of an integrated reporter gene in the absence and presence of anhydrotetracycline, respectively (p < 0.0001), by use of a newly derived minimal promoter based on the core promoter of TgGRA2. In comparison with the SAG1 minimal promoter, use of the GRA2 minimal promoter in stable transfectants has a 23-fold higher Signal to Noise Ratio for EYFP fluorescence in the absence or presence of anhydrotetracycline. We conclude that the performance of TATi for both activation and suppression of transcription can be markedly enhanced by incorporating a GRA2 minimal promoter.
Identifiants
pubmed: 34051228
pii: S0166-6851(21)00035-9
doi: 10.1016/j.molbiopara.2021.111384
pii:
doi:
Substances chimiques
Antigens, Protozoan
0
Bacterial Proteins
0
Gra2 protein, Toxoplasma gondii
0
Luminescent Proteins
0
Membrane Glycoproteins
0
Protozoan Proteins
0
Recombinant Proteins
0
SAG1 antigen, Toxoplasma
0
SAG4 protein, Toxoplasma gondii
0
Tetracyclines
0
yellow fluorescent protein, Bacteria
0
4-epianhydrotetracycline
680VDL31MX
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
111384Informations de copyright
Copyright © 2021 Elsevier B.V. All rights reserved.