Journal of Industrial Microbiology

, Volume 16, Issue 6, pp 342–347

Electrotransformation ofClostridium thermosaccharolyticum

  • T R Klapatch
  • M L Guerinot
  • L R Lynd
Article

DOI: 10.1007/BF01570112

Cite this article as:
Klapatch, T.R., Guerinot, M.L. & Lynd, L.R. Journal of Industrial Microbiology (1996) 16: 342. doi:10.1007/BF01570112

Abstract

Transformation of the thermophileClostridium thermosaccharolyticum ATCC 31960 was achieved using plasmid pCTC1 and electroporation. Evidence supporting transformation was provided by Southern blots, detection of the plasmid in 10 out of 10 erythromycin-resistant clones, retransformation ofE. coli andC. thermosaccharolyticum with plasmid DNA isolated fromC. thermosaccharolyticum, and a proportional relationship between the number of transformants and the amount of DNA added. Transformation efficiencies were very low for plasmid DNA prepared fromE. coli (0.6 transformants mg−1 DNA), although somewhat higher for plasmid DNA prepared fromC. thermosaccharolyticum (52 transformants mg−1 DNA). Transformation-dependent erythromycin resistance indicates that an adenosine methylase gene originating fromEnterococcus faecalis, a mesophile, is expressed inC. thermosaccharolyticum. The plasmid pCTC1 appears to be replicated independently of the chromosome, as indicated by visualization of recovered plasmid on gels, and retransformation using recovered plasmid. pCTC1 is maintained inC. thermosaccharolyticum at both 45 and 60°C. Restriction analysis showed little or no rearrangement occurred upon passage through the thermophile.

Keywords

transformationelectroporationClostridium thermosaccharolyticumthermophile

Copyright information

© Society for Industrial Microbiology 1996

Authors and Affiliations

  • T R Klapatch
    • 1
  • M L Guerinot
    • 1
  • L R Lynd
    • 1
    • 2
  1. 1.Department of Biological SciencesDartmouth CollegeHanoverUSA
  2. 2.Thayer School of EngineeringDartmouth CollegeHanoverUSA
  3. 3.Merck and CoWest PointUSA