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. 1994 Mar;60(3):903-7.
doi: 10.1128/aem.60.3.903-907.1994.

Natural and Electroporation-Mediated Transformation of Methanococcus voltae Protoplasts

G B Patel  1 , J H Nash, B J Agnew, G D Sprott
Affiliations

Natural and Electroporation-Mediated Transformation of Methanococcus voltae Protoplasts

G B Patel et al. Appl Environ Microbiol. 1994 Mar.

Abstract

The lack of high-efficiency transformation systems has severely impeded genetic research on methanogenic members of the kingdom Archaeobacteria. By using protoplasts of Methanococcus voltae and an integration vector, Mip1, previously shown to impart puromycin resistance, we obtained natural transformation frequencies that were about 80-fold higher (705 transformants per mug of transforming DNA) than that reported with whole cells. Electroporation-mediated transformation of M. voltae protoplasts with covalently closed circular Mip1 DNA was possible, but at lower frequencies of ca. 177 transformants per mug of vector DNA. However, a 380-fold improvement (3,417 transformants per mug of DNA) over the frequency of natural transformation with whole cells was achieved by electroporation of protoplasts with linearized DNA. This general approach, of using protoplasts, should allow the transformation of other methanogens, especially those that may be gently converted to protoplasts as a result of their tendency to lyse in hypotonic solutions.

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References

    1. Annu Rev Microbiol. 1992;46:165-91 - PubMed
    1. Appl Environ Microbiol. 1991 Sep;57(9):2762-3 - PubMed
    1. J Bacteriol. 1991 Jun;173(11):3414-8 - PubMed
    1. Mol Gen Genet. 1990 Apr;221(2):273-9 - PubMed
    1. Crit Rev Microbiol. 1989;17(1):53-87 - PubMed

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