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Efficient transfection of the archaebacterium Halobacterium halobium. 1987

S W Cline, and W F Doolittle

We developed an efficient polyethylene glycol-mediated spheroplast transfection method for the extremely halophilic archaebacterium Halobacterium halobium. The 59-kilobase-pair linear phage phi H DNA molecule routinely produced between 5 X 10(6) and 2 X 10(7) transfectants per microgram of DNA. Between 0.5 and 1% of spheroplasts were transfected per microgram of luminal diameter H DNA. Under our conditions, survival and regeneration of H. halobium spheroplasts were also quite efficient, suggesting that this method will be useful for introducing other DNAs into these bacteria.

UI MeSH Term Description Entries
D006217 Halobacterium A genus of HALOBACTERIACEAE whose growth requires a high concentration of salt. Binary fission is by constriction.
D001435 Bacteriophages Viruses whose hosts are bacterial cells. Phages,Bacteriophage,Phage
D013104 Spheroplasts Cells, usually bacteria or yeast, which have partially lost their cell wall, lost their characteristic shape and become round. Spheroplast
D014162 Transfection The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES. Transfections

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