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Journal of Bacteriology, May 1999, p. 2992-3002, Vol. 181, No. 10
Jet Propulsion Laboratory, California
Institute of Technology, Pasadena, California 91109
Received 16 October 1998/Accepted 1 March 1999
Strain PS of Methanococcus voltae (a methanogenic,
anaerobic archaebacterium) was shown to generate spontaneously 4.4-kbp chromosomal DNA fragments that are fully protected from DNase and that,
upon contact with a cell, transform it genetically. This activity,
here called VTA (voltae transfer agent), affects all markers tested: three different auxotrophies (histidine,
purine, and cobalamin) and resistance to BES (2-bromoethanesulfonate, an inhibitor of methanogenesis). VTA was most effectively prepared by
culture filtration. This process disrupted a fraction of the M. voltae cells (which have only an S-layer covering their
cytoplasmic membrane). VTA was rapidly inactivated upon
storage. VTA particles were present in cultures at concentrations of
approximately two per cell. Gene transfer activity varied from a
minimum of 2 × 10
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Transduction-Like Gene Transfer in the Methanogen
Methanococcus voltae
5 (BES resistance) to a maximum of
10
3 (histidine independence) per donor cell. Very little
VTA was found free in culture supernatants. The phenomenon is
functionally similar to generalized transduction, but there is no
evidence, for the time being, of intrinsically viral (i.e., containing
a complete viral genome) particles. Consideration of VTA DNA size makes the existence of such viral particles unlikely. If they exist,
they must be relatively few in number;perhaps they differ from
VTA particles in size and other properties and thus escaped detection.
Digestion of VTA DNA with the AluI restriction enzyme suggests that it is a random sample of the bacterial DNA, except for a
0.9-kbp sequence which is amplified relative to the rest of the
bacterial chromosome. A VTA-sized DNA fraction was demonstrated in a
few other isolates of M. voltae.
*
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