A dual expression plasmid with Microcin B17 compatible with both prokaryotic and mammalian systems

Area of Science:

• Molecular Biology
• Biotechnology

Background:

• Proteic plasmid addiction systems, like control of cell death (Ccd), are vital for efficient plasmid DNA recombination.
• The CcdB toxin, a common choice, is lengthy (309 bp), driving the search for shorter peptide toxins.

Purpose of the Study:

• To evaluate microcin B17 (MccB17), a 43-amino acid peptide, for DNA recombination in the pgMAX-II dual expression plasmid.
• To assess the efficiency of MccB17 in facilitating gene cloning and dual expression in prokaryotic and mammalian systems.

Main Methods:

• Utilized the pgMAX-II dual expression plasmid system.
• Incorporated the α-peptide gene from lacZ for α-complementation.
• Tested MccB17's efficacy in promoting DNA recombination and gene expression (DsRed) in *E. coli* and mammalian cells.

Main Results:

• The pgMAX-II/MccB17 plasmid demonstrated highly efficient cloning of external DNA via α-complementation.
• Successful expression of the DsRed fluorescent protein gene was achieved in both *E. coli* and mammalian cells.
• MccB17 showed comparable potential for DNA recombination to CcdB.

Conclusions:

• The novel pgMAX-II/MccB17 plasmid enables efficient and straightforward subcloning of external genes.
• This system supports dual expression in both prokaryotic (*E. coli*) and mammalian cells, indicating broad applicability.
• MccB17 offers a viable, shorter alternative for plasmid addiction systems in versatile dual-expression vectors.