Plasmids : a practical approach


Plasmids : a practical approach

edited by K.G. Hardy

(The practical approach series)

IRL Press at Oxford University Press, c1993

2nd ed

  • pbk.

大学図書館所蔵 件 / 17



Includes bibliographical references and index



pbk. ISBN 9780199634446


This book is designed to provide research workers with complete and detailed protocols for studying bacterial plasmids and for using both plasmids and phagemids as vectors. Techniques for studying fundamental aspects of plasmids, such as replication, are included as well as methods for using plasmid vectors in important groups of bacteria. Also described are protocols for using phagemids, which are extremely useful tools for genetic engineering.


  • 1. Plasmid replication and maintenance
  • 2. Plasmids from gram-positive bacteria
  • 3. Lactococcal plasmid vectors
  • 4. Virulence plasmids
  • 5. Plasmids in Agrobacterium tumefaciens and Rhodococcus fascians
  • 6. Streptomyces plasmid vectors
  • 7. Use of -plasmid composite vectors for expression cDNA cloning
  • 8. Phagemids and other hybrid vectors
  • 9. Extrachromosomal cloning vectors of Saccharomyces cerevisiae
  • Appendices

ISBN 9780199634453


This work is designed to provide research workers with detailed protocols for studying bacterial plasmids, and for using both plasmids and phagemids as vectors. The techniques for studying plasmid replication and maintenance are described in detail, as are fundamental methods for purifying plasmids and for introducing them into cells. Also included are protocols for mutagenesis, and for analyzing plasmid-encoded products. Other chapters focus on techniques for studying plasmids and for using plasmid vectors in particularly important groups of bacteria - animal and plant pathogens harbouring virulence plasmids, streptomyces and lactococci. Plasmid composite vectors and phagemids of various kinds are useful for expression, cDNA cloning and other genetic manipulations. The protocols for using them, as well as important yeast shuttle vectors are described in detail.


  • Part 1 Plasmid replication and maintenance, K. Nordstroem: single-cell resistance (SCR) to antibiotics
  • determination of plasmid copy numbers
  • partition and other maintenance functions
  • incompatibility
  • basic replicons
  • origins (ori) of replication
  • rep functions
  • identification cis/trans activity
  • replicative intermediates
  • timing intermediates
  • timing of replication
  • invitro replication
  • visualization of plasmids. Part 2 Plasmids from gram-positive bacteria, J.C. Alonso and Manuel Espinosa: classification
  • isolation of plasmid DNA
  • host range and plasmid tranfer
  • plasmid replication modes. Part 3 Lactococcal plasmid vectors, K.J. Leenhouts and G. Venema: Lactococcal vector systems
  • special-purpose vectors. Part 4 Virulence plasmids, Marcelo E. Tomalsky et al: isolation of plasmids
  • curing of plasmids
  • mutagenesis
  • fast and small electrophoresis-sequencing gels
  • comparison of helical stability of DNA fragments
  • identification of plasmid-encoded products
  • RNA extraction and analysis. Part 5 Plasmids in agrobacterium tumefaciens and rhodococcus fascians, Jan Desomer et al: identification of endogenous plasmids by small-scale isolation procedures
  • large-scale purification methods
  • introductioon of recombinant plasmids
  • analysis of plasmid-encoded functions. Part 6 Streptomyces plasmid vectors, W. Wohlleben and G. Muth: autonomously replicating streptomyces vectors
  • integrating vectors
  • gene disruption/replacement techniques using temperature-sensitive and non-replicative plasmids. Part 7 Use of lambda-plasmid composite vectors for expression cDNA cloning, Toru Miki and Stuart A. Aaronson: expression cloning using unidirectional cDNA libraries
  • generation of cDNA libraries by the ADC method
  • analysis of cloned cDNAs
  • applications. Part 8 Phagemids and other hybrid vectors, Michelle A. Alting-Mees et al: early vectors
  • new applications
  • phagemids and lambda-zedAP cloning. Part 9 Extrachromosonal cloning vectors of saccharomyces cerevisiae, Robert S. Sikorski: basic cloning vectors of saccharomyces cerevisiae
  • multifunctional cloning vectors of saccharomyces cerevisiae
  • yeast host strain
  • introduction of DNA into saccharomyces cerevisiae. Appendix: supplies of specialist items.

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