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Superiority of the PCR-based Approach for Cloning the Acetate Kinase Gene of Clostridium Thermocellum
Industrial Microbiology and Biotechnology
  • G. Ozcengiz
  • J-H Kim
  • E. Ozcengiz
  • David J. Westenberg, Missouri University of Science and Technology
  • W. R. Lin
  • L. R. Lynd
  • A. L. Demain
Cloning of Clostridium thermocellum acetate kinase (ack) and/or phosphotransacetylase (pta) genes in Escherichia coli by functional complementation of ack and/or pta mutants was complicated by an alternative acetate assimilation pathway involving acetyl-CoA synthetase (ACS). In addition to the problems encountered with the complementation approach, cloning of these genes was not readily achieved using heterologous probing with corresponding genes from Escherichia coli and Methanosarcina thermophila due to the lack of sufficient homology. The use of a PCR-based approach, on the other hand, yielded a specific C. Thermocellum gene fragment which showed significant sequence identity to the ack gene for which primers were designed. The subcloned ack fragment was then successfully used as a probe for the isolation of the corresponding gene and restriction analysis of that region.
Biological Sciences
J. William Fulbright Scholarship
Document Type
Article - Journal
Document Version
File Type
© 1998 Springer Verlag, All rights reserved.
Publication Date
Citation Information
G. Ozcengiz, J-H Kim, E. Ozcengiz, David J. Westenberg, et al.. "Superiority of the PCR-based Approach for Cloning the Acetate Kinase Gene of Clostridium Thermocellum" Industrial Microbiology and Biotechnology (1998)
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