Stable plasmid expression vector for bacteria
View Patent ↗The invention provides compositions and methods for stable plasmid maintenance and protein expression in bacteria. Further provided are compositions and methods for promoting competence in bacteria that are otherwise not transformable.
1. An isolated recombinant plasmid expression vector comprising a polynucleotide encoding a prokaryote-derived actin like protein (ALP) having at least 90% identity to the polypeptide sequence of SEQ ID NO:1, wherein the ALP confers stability on a mobile genetic element when the ALP is expressed in a prokaryotic cell having a mobile genetic element.
2. The vector of claim 1 , wherein the vector encodes an ALP7.
3. The vector according to claim 1 , wherein the ALP comprises: D at the residue corresponding to human beta actin residue 11, G at the residue corresponding to human beta actin residue 13, E or Q at the residue corresponding to human beta actin residue 137, D at the residue corresponding to human beta actin residue 154, and G at the residue corresponding to human beta actin residue 156, when optimally aligned with the polypeptide sequence of human beta actin.
4. The vector of claim 1 , further comprising a polynucleotide encoding a polypeptide sequence having at least 90% identity to SEQ ID NO:411.
5. The vector of claim 1 , further comprising a polynucleotide sequence that encodes a heterologous polypeptide.
6. The vector of claim 5 , wherein the heterologous polypeptide is competence factor K (comK).
7. The vector of claim 6 , further comprising an origin of transfer.
8. An isolated bacteria comprising the plasmid expression vector of claim 1 .
9. The bacteria of claim 8 , wherein the bacteria is a Bacillus or Escherichia strain.