Molecular biological methods for Bacillus


Molecular biological methods for Bacillus

edited by Colin R. Harwood and Simon M. Cutting ; with contributions by R. Chambert ... [et al.]

(Modern microbiological methods)

Wiley, c1990

大学図書館所蔵 件 / 14



"A Wiley-Interscience publication."

Includes bibliographical references

Includes index



This series aims to identify specialist areas in microbiology and provide up-to-date methodological handbooks to aid microbiologists at the laboratory bench. The books are directed primarily toward active research workers but are structured so as to serve as an introduction within a speciality for graduate students and scientists entering microbiology from related disciplines. This book focuses on the growth, maintenance and techniques associated with Bacillus. It deals with genetic analysis and then discusses the use of plasmids, transposons and integrational vectors and gene cloning techniques for Bacillus. The book goes on to consider the measurement of gene expression in Bacillus and the mechanisms of DNA repair and replication in this organism. The Bacillus cell envelope and secretion are examined and sporulation is discussed. Finally the book discusses the culturing and maintenance of bacteriophages and provides physical and genetic maps of Bacillus subtilis. Methods and references are given at the end of each section.


  • Part 1 Growth maintenance and general techniques, C.R.Harwood and A.R.Archibald: the genus Bacillus
  • growth of Bacillus
  • maintenance and shipping of strains
  • cell permeabilization and breakage
  • radiolabelling of macromolecules. Part 2 Genetic analysis, S.M.Cutting and P.B.Vander Horn: mutagenesis
  • transformation
  • PBSI generalized transduction
  • protoplast techniques
  • selection and screening of recombinants
  • genetic mapping
  • complementation analysis. Part 3 Plasmids, S.Bron: plasmids used in Bacillus
  • isolation of plasmid DNA
  • plasmid DNA transfer systems for Bacillus
  • plasmid replication in Bacillus
  • plasmid instability in Bacillus subtilis
  • efficient cloning systems for Bacillus subtilis based on pTA1060 and pAMbeta1. Part 4 Gene cloning techniques, J.Errington: cloning vectors
  • cloning systems
  • special purpose cloning vectors
  • special purpose hosts. Part 5 Use of transposons and integrational vectors mutagenesis and construction of gene fusionson Bacillus species, P.Youngman: insertional mutagenesis with Tn917
  • insertional mutagenesis with integrational vectors
  • using transposons or integrational vectors to tag specific regions of the chromosome
  • cloning of chromosomal sequences adjacent to transposon insertions or plasmid integrations
  • construction of lacZ fusions using transposons, integrational vectors and temperate phages
  • information concerning vectors, strains and protocols. Part 6 Measuring gene expression in Bacillus, C.P.Moran: transcription products
  • translation products
  • transcription in vitro. Part 7 DNA repair and replication in Bacillus, R.E.Yasbin et al: DNA repair mechanisms
  • DNA repair in Bacillus
  • DNA replication. Part 8 The Bacillus cell envelope and secretion, C.R.Harwood et al: structure and function of the cell envelope
  • synthesis and maturation of the Bacillus cell wall
  • protein secretion
  • electron microscopy for bacterial cells
  • electron microscope methodologies. Part 9 Sporulation, W.L.Nicholson and P.Setlow: induction of sporulation
  • sporulation - specific marker events
  • isolation and analysis of small, acid-soluble spore proteins
  • extraction and analysis of spore coat proteins
  • sequential gene expression
  • compartmentalization
  • purification of spores
  • storage of purified spores
  • isolation and characterization of germination mutants
  • isolation of spore growth mutants
  • isolation of sporulation mutants. Part 10 Bacteriophages, H.E.Hemphill: host bacteria for growing B. subtilis phages
  • media for culturing and maintaining B. subtilis phages
  • culturing and maintaining bacteriophage stocks
  • titering B. subtilis phages
  • batch culturing of bacteriophages
  • lysogens of temperate B. subtilis bacteriophages
  • phage isolation and purification
  • isolation of bacteriophage
  • bacteriophage mutants. (Part contents).

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