Golden Gate Cloning of Expression Plasmids for Synthetic Small RNAs in Bacteria.
Methods Mol Biol
; 2850: 307-328, 2025.
Article
in En
| MEDLINE
| ID: mdl-39363079
ABSTRACT
Bacterial small RNAs (sRNAs) are well known for their ability to modulate gene expression at the post-transcriptional level. Their rather simple and modular organization provides the user with defined building blocks for synthetic biology approaches. In this chapter, we introduce a plasmid series for Escherichia coli and describe protocols for fast and efficient construction of synthetic sRNA expression plasmids based on Golden Gate assembly. In addition, we present the G-GArden tool, which assists with the design of oligodeoxynucleotides and overhangs for scarless assembly strategies. We propose that the presented procedures are suitable for many applications in different bacteria, which are related to E. coli and beyond.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Plasmids
/
RNA, Bacterial
/
Cloning, Molecular
/
Escherichia coli
Language:
En
Journal:
Methods Mol Biol
Journal subject:
BIOLOGIA MOLECULAR
Year:
2025
Document type:
Article
Affiliation country:
Germany
Country of publication:
United States