A standardized set of pNX vectors for enhanced soluble expression of recombinant proteins in E. coli using small fusion tags.

ABSTRACT:

Background

The production of recombinant proteins in Escherichia coli (E. coli) is often hampered by the formation of inclusion bodies. While fusion tags can enhance solubility, existing systems are hampered by a lack of standardization, with tags scattered across disparate plasmid backbones and inconsistent cloning sites, complicating parallel screening.

Results

To address this, we constructed a standardized series of expression vectors, termed pNX, by incorporating nine small fusion tags (SUMO, LD, ACP, BCCP, GB1, Fh8, SmbP, TolA, and TrxA) into a uniform pET-28b backbone. Each pNX vector features an identical configuration: a T7 promoter, an N-terminal fusion tag, a synthetic linker, a TEV protease cleavage site, and a multiple cloning site (MCS) flanked by dual 6×His tag

SUBMITTER: Luo LZ 

PROVIDER: S-EPMC12825285 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature