pKNOCK-Bla-ermG plasmid suicide vector自殺質(zhì)粒載體

pKNOCK-bla-ermGa/b suicide vectors (Structure 16: 1-11). Base suicide vectors contain ermG for selection on erythromycin inBacteroides spp. and bla for selection on ampicillin in E. coli. Vectors are derivatives of pKNOCK-cm (Biotechniques 26(5): 824-828).The complete nucleotide sequence has been deduced. The pBR325 cat gene was removed from pKNOCK-cm by MluI digestion and replacedwith a PCR-amplified, bla-ermG-containing fragment from pGERM (Methods 20: 35-46). The added bla-ermG fragment, with MluI ends,was inserted into the pKNOCK-cm MluI sites in both possible orientations (“a” and “b” designation). Select restriction sites are shown alongwith the multiple cloning site (MCS), R6K replication origin (oriR6K) and the RP4 transfer origin (RP4 oriT). In general, the “b” version wasused for downstream constructions.

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