Avoidance of reporter assay distortions from fused dual reporters.
RNA
; 23(8): 1285-1289, 2017 08.
Article
en En
| MEDLINE
| ID: mdl-28442579
ABSTRACT
Positioning test sequences between fused reporters permits monitoring of both translation levels and framing, before and after the test sequence. Many studies, including those on recoding such as productive ribosomal frameshifting and stop codon readthrough, use distinguishable luciferases or fluorescent proteins as reporters. Occasional distortions, due to test sequence product interference with the individual reporter activities or stabilities, are here shown to be avoidable by the introduction of tandem StopGo sequences (2A) flanking the test sequence. Using this new vector system (pSGDluc), we provide evidence for the use of a 3' stem-loop stimulator for ACP2 readthrough, but failed to detect the reported VEGFA readthrough.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Genes Reporteros
/
Codón de Terminación
/
Sistema de Lectura Ribosómico
/
Luciferasas
/
Proteínas Luminiscentes
Límite:
Humans
Idioma:
En
Revista:
RNA
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2017
Tipo del documento:
Article
País de afiliación:
Irlanda