The pNZ8148 vector contains an origin of
replication (ORI), the gene for the resistance to chloramphenicol, two genes
for the replication proteins repA and repC, the nisin-inducible promoter (P
nisA), and the transcription terminator (T). The gene, tagged with a Strep-tag
II (STREP) followed by a stop codon (*), can be inserted between the necessary
NcoI site and another endonuclease site from the multicloning site (MCS) such
as PstI, SphI, KpnI, SpeI, XbaI, SacI and HindIII. The replicons of the vectors
pNZ8008, pNZ8148, pNZ8149 and pNZ8150 are identical and came originally from
the Lactococcus lactis plasmid pSH71. However, this replicon has a broad host
range. Plasmids with this replicon can replicate in many Gram-positive
bacteria, such as Lactobacillus plantarum and Streptococcus thermophilus. pNZ8148 – In this vector
the nisA promoter is followed by an NcoI site for translational fusions at the
ATG. It contains a terminator after the MCS. Sequence adaptation for cloning in
NcoI can result in a change in the second amino acid of a protein (Mierau and
Kleerebezem, 2005).