HR: 0800h
AN: V41B-1371    [Abstracts]
TI: Thermo-resistance Acquisition of A Mesophilic Bacterium with The Aid of Vector Particles Originating from Thermophiles
AU: * Sugitate, T
EM: sugitate@ca2.so-net.ne.jp
AF: Biology, Int'l Christian Univ, 10-2, Osawa 3-chome, Mitaka-shi, Tokyo, 181-8585 Japan
AU: Inaba, N
EM: chiura@icu.ac.jp
AF: Biology, Int'l Christian Univ, 10-2, Osawa 3-chome, Mitaka-shi, Tokyo, 181-8585 Japan
AU: Kurusu, Y
EM: krsy@mx.ibaraki.ac.jp
AF: Fac Agric, Ibaraki Univ, Chuo 3-21-1, Ami-machi, Inashiki-gun, Ibaraki, 300-0393 Japan
AU: Hoaki, T
EM: toshihiro.hoaki@sakura.taisei.co.jp
AF: Civil Enging Res Inst, Technology Centre, Taisei Corp, 344-1, Nase-cho, Totsuka-ku, Yokohama, 245-0051 Japan
AU: Chiura, H X
EM: chiura@icu.ac.jp
AF: Biology, Int'l Christian Univ, 10-2, Osawa 3-chome, Mitaka-shi, Tokyo, 181-8585 Japan
AB: The present study was aimed to examine whether virus-like particles (VLPs) would be able to transfer and express the thermo-resistance gene of thermophilic microbes in the mesophilic auxotrophic {\it Escherichia coli} AB1157 mutant. A hyper-thermophilic archaea, {\it Thermococcus kodakaraensis} B41, that was isolated from the Suiyo Seamount APSK06 boring core, released particles (KD-VLPs) during culture. Transduction towards recipient {\it E. coli} AB1157 was carried out using KD-VLP as the gene transfer mediator, in order to examine the lethal effect and thermo-resistant gene transfer capability of the particle. The colony forming ability of the cells was examined in 7 $%$ of gelrite supplemented-LB plates (LB-gelrite plates) at 50, 56, and 70 $\deg$C. Regardless of UV irradiation, KD-VLP showed a reduced efficiency of plating (EOP) of recipient viable cell population to ca 65 $%$. Four colonies were formed in LB-gelrite plates at 50 $\deg$C, which were named as KD-E-Trans, and the gene transfer frequency was estimated to be 5.12 $\times$ 10$^{-8}$ cfu/particle. Obtained KD-E-Trans was cultured in LB liquid medium employing the same high temperature conditions. The cells grew 1.6 $\sim$ 6 fold of the inocula in 13 days at all the examined temperatures, and the generation time of the transductants were as follows: ca 28 hours at 50 $\deg$C, ca 73 hours at 56 $\deg$C, ca 266 hours at 70 $\deg$C. Thus, the gene transfer of thermo-resistance to mesophilic {\it E. coli} from another Domain with the aid of KD-VLPs was demonstrated.
DE: 4832 Hydrothermal systems
DE: 4840 Microbiology
DE: 4800 OCEANOGRAPHY: BIOLOGICAL AND CHEMICAL
DE: 4803 Bacteria
SC: Volcanology, Geochemistry, Petrology [V]
MN: 2004 AGU Fall Meeting