Isolation and Characterisation of Broad-Host Range Gene Transporter Particles from Geo-Thermal Vent of the Toyoha Mine

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Virus-like particles (VLPs) were collected from geothermal vent water samples in the drift-way at Toyoha Mine, Hokkaido, Japan (-500 m level, 63.5℃) whose VLP and bacterial abundance was (No/ml±SD, n: 500), VLP: 9.60±0.29×10^8 and bacteria: 3.61±0.14×10^6. VLPs ranged in diameter from 30 to 320 nm, and the major size distribution (ca 62%) was 83.3±3.3 nm (n: 843). Ultrafiltration followed by CsCl density equilibrium ultracentrifugation gave purified TY-VLPs: 6.64×1 0^<13>. Regardless of UV treatment, TY-VLP reduced the efficiency of plating to 68.6-83.4% at a multiplicity of infection of ca 0.3 on Escherichia coli AB1157. Generalised transduction was observed on E. coli AB1157 with a frequency between 10^<-4> and 10^<-5> cells/particle using TY-VLPs without UV-treatment. The growth of generated E. coli transductants (TY-E-trans) was compared to that of an E. coli transductant (ST-E-trans) generated by Aquificales originating VLP (Chiura, 2002). The extent of the maximum growth of both transductants was ca 40% of the parental E. coli used as a recipient. TY-E-trans acquired "budding-like" particle productivity, which has been demonstrated for ST-E-trans. ST-E-trans produced five different size particles, whose DNA content ranged between 291.6 and 382.0 kb, and TY-E-trans produced ten different size particles between 68.5 and 190.2 kb, respectively.
日本微生物生態学会の論文