ANALYSIS OF THE ROLES OF THE cwlD OPERON PRODUCTS DURING SPORULATION IN BACILLUS SUBTILIS
نویسندگان
چکیده
ii ACKNOWLEDGEMENTS iv LIST OF FIGURES v LIST OF TABLES vii CHAPTER I. REVIEW OF THE LITERATURE Stages of sporulation 1 Resistance properties of bacterial endospores 3 Spore peptidoglycan 3 The CwlD protein 5 Phenotypic characteristics of the cwlD mutant 5 Synthesis of muramic-δ-lactam 6 The cwlD operon 7 CHAPTER II. THE BIOCHEMICAL ANALYSIS OF CWLD Materials and Methods 11 Construction of strains and plasmids 11 E.coli strain construction and growth media 12 Induction of CwlD 14 Separation of protein fractions 14 Purification of CwlD with metal affinity chromatography 14 Purification of CwlD with anion exchange chromatography 15 Purification of Bacillus subtilis vegetative peptidoglycan 15 Purification of E.coli peptidoglycan 16 Testing activity of purified CwlD in vitro 17 Detecting CwlD activity in vivo 17 Production and purification of polyclonal anti-CwlD 18 antibodies Results from the Biochemical Analysis of CwlD 19 Strains analyzed 19 Induction of CwlD 19
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