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Cloning DNA from Rhizobium Meliloti Using a New Broad Host Range, Binary Vehicle System

  • G. Ditta
  • S. Stanfield
  • D. Corbin
  • D. R. Helinski

Abstract

Molecular cloning techniques now permit the dissection of DNA to almost any desired degree in order to study both structure and function of individual genes. An important strategy for analyzing cloned DNAs involves reintroducing them into their host of origin under various conditions designed to affect gene function. This procedure is greatly facilitated if the primary cloning vector is capable of autonomous replication in multiple hosts. Double vectors, capable of replication in more than one host, have been developed for use in Escherichia coli/yeast (Struhl et al., 1979) and Bacillus subtilis/Staphylococcus aureus (Lofdahl et al., 1978). There are many other organisms of medical, agricultural, and economic importance, however, for which no such dual purpose vectors exits. We have therefore developed a broad host range cloning system for gram-negative bacteria from the naturally-occurring drug resistance plasmid RK2. This system, whose possible development was first suggested some time ago (Meyer et al., 1977), has proved exceedingly valuable to our laboratory in working with the nitrogen-fixing alfalfa symbiont, Rhizobium meliloti.

Keywords

Gene Bank Broad Host Range Helper Plasmid Transfer Frequency Acinetobacter Calcoaceticus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1981

Authors and Affiliations

  • G. Ditta
    • 1
  • S. Stanfield
    • 1
  • D. Corbin
    • 1
  • D. R. Helinski
    • 1
  1. 1.University of California, San DiegoLa JollaUSA

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