Cloning of pduJ for High-Level Expression . 60 Purification of the PduA Protein. 61 Antiserum Preparation . 61 W estern Blots. 62 Electron Microscopy. 63 Construction of a Nonpolar pduA Deletion . 64 DNA Sequencing and Analysis . 64
    Results . 65
        Purification of Recombinant His6-PduA Protein . 65 Preparation of PduA-Specific Antiserum . 68 Localization of PduA by Immunoelectron Microscopy. 70 Localization of Diol Dehydratase in apduA Mutant . 70 The pduA Gene is Required for the Formation of Polyhedral Organelles .72 Co-Localization of the PduA Protein and Diol Dehydratase . 72
        Complementation of the Polyhedral Organelle Minus Phenotype of a zlpduA
          Mutant Strain. 74
        Strains with pduA Mutations Show a Period of "Interrupted" Growth when
          Cultured on PD/CN-B12 Minimal Medium . 76
        CO2 Supplementation Does not Correct the "Interrupted" Growth Phenotype
          of apduA M utant  Strain  . .78
        Effects of PD Concentration on the Growth Rates of Wild Type and pduA
          Mutant Strains. 78
        Effects of CN-B312 Concentration on the Growth Rates of Wild Type and pduA
          Null Mutant Strains. 83
    Discussion . 85

4 PURIFICATION ANT) IDENTIFICATION OF THlE MAJOR PROTEINS OF
    POLYHEDRAL ORGANELLES INVOLVED IN COENZYME B12-DEPEND)ENT
    DEGRADATION OF 1,2-PROPANEDIOL IN SALMONELLA ENTERICA
    SERO VAR TYPHIMURIUM LT2 .89

    Introduction. 89 Materials and Methods. 92
        Chemicals and Reagents . 92 Organelle Purification . 92 General Protein and Molecular Methods . 94 Electron Microscopy. 94 W estern Blots. 94 Enzyme Assays . 94 Glycoprotein  Staining. 95 N-terminal Sequencing. 95
        Matrix Assisted Laser Desorption Ionization-Time of Flight Mass
          Spectrometry (MALDI-TOF MS) . 96
        Peptide Mass Fingerprinting. 97 Densitometry Analysis. 98
    Results . 98
        Purification of Polyhedral Organelles . 98