Analysis of Orientia tsutsugamushi promoter activity.
Animals
Antigens, Bacterial
/ genetics
Bacterial Outer Membrane Proteins
/ genetics
Bacterial Proteins
/ genetics
Cell Line
Green Fluorescent Proteins
/ genetics
Haplorhini
HeLa Cells
Humans
Orientia tsutsugamushi
/ genetics
Promoter Regions, Genetic
Rickettsia Infections
/ microbiology
Scrub Typhus
/ microbiology
THP-1 Cells
Orientia tsutsugamushi
Rickettsia
Rickettsiales
promoter
scrub typhus
Journal
Pathogens and disease
ISSN: 2049-632X
Titre abrégé: Pathog Dis
Pays: United States
ID NLM: 101595366
Informations de publication
Date de publication:
23 09 2021
23 09 2021
Historique:
received:
23
07
2021
accepted:
10
09
2021
pubmed:
14
9
2021
medline:
4
2
2022
entrez:
13
9
2021
Statut:
ppublish
Résumé
Orientia tsutsugamushi is an obligate intracellular bacterium that causes scrub typhus, a potentially fatal rickettsiosis, and for which no genetic tools exist. Critical to addressing this technical gap is to identify promoters for driving expression of antibiotic resistance and fluorescence reporter genes in O. tsutsugamushi. Such promoters would need to be highly conserved among strains, expressed throughout infection, and exhibit strong activity. We examined the untranslated regions upstream of O. tsutsugamushi genes encoding outer membrane protein A (ompA), 22-kDa type-specific antigen (tsa22) and tsa56. The bacterium transcribed all three during infection of monocytic, endothelial and epithelial cells. Examination of the upstream noncoding regions revealed putative ribosome binding sites, one set of predicted -10 and -35 sequences for ompA and two sets of -10 and -35 sequences for tsa22 and tsa56. Comparison of these regions among geographically diverse O. tsutsugamushi patient isolates revealed nucleotide identities ranging from 84.8 to 100.0%. Upon examination of the candidates for the ability to drive green fluorescence protein expression in Escherichia coli, varying activities were observed with one of the tsa22 promoters being the strongest. Identification and validation of O. tsutsugamushi promoters is an initial key step toward genetically manipulating this important pathogen.
Identifiants
pubmed: 34515306
pii: 6369350
doi: 10.1093/femspd/ftab044
pmc: PMC8460164
pii:
doi:
Substances chimiques
Antigens, Bacterial
0
Bacterial Outer Membrane Proteins
0
Bacterial Proteins
0
Green Fluorescent Proteins
147336-22-9
OMPA outer membrane proteins
149024-69-1
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NIAID NIH HHS
ID : R01 AI123346
Pays : United States
Organisme : NIAID NIH HHS
ID : R21 AI152513
Pays : United States
Informations de copyright
© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
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