Identification of Cryptic Promoter Activity in cDNA Sequences Corresponding to PRRSV 5' Untranslated Region and Transcription Regulatory Sequences.
Viruses
; 14(2)2022 02 15.
Article
in En
| MEDLINE
| ID: mdl-35215993
ABSTRACT
To investigate the role of PRRSV nonstructural proteins (nsps) in viral RNA replication and transcription, we generated a cDNA clone of PRRSV strain NCV1 carrying the nanoluciferase (nluc) gene under the control of the transcription regulatory sequence 6 (TRS6) designated as pNCV1-Nluc. Cells transfected with the pNCV1-Nluc DNA plasmid produced an infectious virus and high levels of luciferase activity. Interestingly, cells transfected with mutant pNCV1-Nluc constructs carrying deletions in nsp7 or nsp9 regions also exhibited luciferase activity, although no infectious virus was produced. Further investigation revealed that the cDNA sequences corresponding to the PRRSV 5' untranslated region (UTR) and TRS, when cloned upstream of the reporter gene nluc, were able to drive the expression of the reporter genes in the transfected cells. Luciferase signals from cells transfected with a reporter plasmid carrying PRRSV 5' UTR or TRS sequences upstream of nluc were in the range of 6- to 10-fold higher compared to cells transfected with an empty plasmid carrying nluc only. The results suggest that PRRSV 5' UTR and TRS-B in their cDNA forms possess cryptic eukaryotic promoter activity.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Promoter Regions, Genetic
/
DNA, Complementary
/
Porcine respiratory and reproductive syndrome virus
/
5' Untranslated Regions
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
Viruses
Year:
2022
Document type:
Article
Affiliation country: