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Literature DB >> 2768194

Transformation of the archaebacterium Halobacterium volcanii with genomic DNA.

S W Cline¹, L C Schalkwyk, W F Doolittle.

Abstract

We describe optimization of a transformation system for the halophilic archaebacterium Halobacterium volcanii. Transformation of spheroplasts in the presence of polyethylene glycol permits the uptake and expression of high-molecular-weight linear fragments of genomic DNA as well as plasmid or bacteriophage DNA. Transformations can be performed with either fresh or frozen cell preparations. Auxotrophic mutants were transformed to prototrophy with genomic DNA from wild-type cells with efficiencies of 5 x 10(4)/micrograms of DNA and frequencies of 8 x 10(-5) per regenerated spheroplast. The overall efficiency of transformation with genomic DNA implies that genetic recombination is an efficient process in H. volcanii.

Entities: Chemical Species

Mesh：

Substances：
DNA, Bacterial

Year: 1989 PMID： 2768194 PMCID： PMC210307 DOI： 10.1128/jb.171.9.4987-4991.1989

Source DB: PubMed Journal: J Bacteriol ISSN： 0021-9193 Impact factor: 3.490

17 in total

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7. Isolation and partial characterization of plasmids found in three Halobacterium volcanii isolates.

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4. Improved growth and morphological plasticity of Haloferax volcanii.

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5. Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance.

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7. Characterization of a gene involved in histidine biosynthesis in Halobacterium (Haloferax) volcanii: isolation and rapid mapping by transformation of an auxotroph with cosmid DNA.

Authors: R K Conover; W F Doolittle
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