Next, a gene cassette with the pMV158-mobM gene was amplified by PCR and cloned into pCR2.1-TOPO. The resulting plasmid (PCR TOPO mob) contains the mobM gene (which has two restriction sites: Xhol and Smal). The pCR TOPO mob plasmid was digested with Xhol and Smal enzymes in order to isolate the mobM fragment. This digestion causes
Next, a gene cassette with the pMV158-mobM gene was amplified by PCR and cloned into pCR2.1-TOPO. The resulting plasmid (PCR TOPO mob) contains the mobM gene (which has two restriction sites: Xhol and Smal). The pCR TOPO mob plasmid was digested with Xhol and Smal enzymes in order to isolate the mobM fragment. This digestion causes