Cloning with plasmid pBR322 BamHI site APR pBR322 TcR Insertion of foreign DNA BamHI sites causes inactivation of the BamHl digestion tetracycline resistance gene Foreign DNA BamHI (insertional inactivation) digestion Clal EcoRI Hind IIl DNA ligase Ampicillin BamHI Recyclized pBR322; Hybrid containing resistance no foreign DNA foreign DNA s PstI Sal I ApR TCR ApR Transformation of E.coli Transformants Transformants resistant to both resistant to ampicillin ampicillin and tetracycline but sensitive to tetracycline -Tetracycline resistance Figure 10-37 Brock Biology of Microorganisms 11/e Origin of DNA 2006 Pearson Prentice Hall,Inc. replicationChen Feng, Lecture of Microbiology Cloning with plasmid pBR322 Insertion of foreign DNA causes inactivation of the tetracycline resistance gene (insertional inactivation)