Influence of the expression vector and its elements on recombinant human prolactin synthesis in Escherichia coli; co-directional orientation of replication and transcription is highly critical
J Microbiol Methods, v. 191, 106340, dez. 2021
Article
en En
| SES-SP, SESSP-IBPROD, SES-SP
| ID: bud-4055
Biblioteca responsable:
BR78.1
ABSTRACT
The aim of the present work was to define a bacterial expression system that is particularly efficient for the synthesis of recombinant human prolactin (hPRL). In previous work, based on experiments that were basically carried out in parallel with the present ones, the synthesis of rec-hPRL by the p1813-hPRL vector in E. coli HB2151 was >500 mg/L, while it was much lower here (2.5–4-fold), in the RB791 and RRI strains. The highest positive influence on rec-hPRL synthesis was due to the transcription-replication co-orientation of hPRL cDNA and the ori/antibiotic resistance gene, responsible for up to a ~ 5–6-fold higher expression yield. In conclusion, this work confirmed that each bacterial strain of E. coli has a genetic background that can allow a different level of heterologous protein synthesis. The individual study of each element indicated that its action critically depends on the reading orientation in which it is located inside the vector co-directional orientation of replication and transcription, in fact, greatly increased the level of rec-hPRL expression.
Texto completo:
1
Colección:
06-national
/
BR
Base de datos:
SES-SP
/
SESSP-IBPROD
Idioma:
En
Revista:
J Microbiol Methods
Año:
2021
Tipo del documento:
Article