TY - GEN
T1 - GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks
AU - Yamaji, Kazuaki
AU - Kanai, Tamotsu
AU - Nomura, Shin Ichiro M.
AU - Akiyoshi, Kazunari
AU - Negishi, Makiko
AU - Atomi, Haruyuki
AU - Yoshikawa, Kenichi
AU - Imanaka, Tadayuki
PY - 2009/12/1
Y1 - 2009/12/1
N2 - The possibilities of an in vitro translation system, based on cell components of the hyperthermophilic archaeon, Thermococcus kodakaraensis, to utilize at a lower temperature range were examined. Green fluorescence protein (GFP) was used as the reporter protein. Through optimization of several experimental conditions, the yield of active GFP production at 40°C increased 5-fold. The optimized system was encapsulated in liposome made by passing through the oil/water interfaces. The prepared liposomes were incubated at 40°C for 90 min, and fluorescence was examined by laser confocal microscopy. A significant increase in average fluorescence intensity was observed in liposomes with GFP mRNA, indicating that the T. kodakaraensis in vitro translation system is applicable for protein production within giant liposomes. These artificial cell models should provide the means to reconstitute various cell functions from a synthetic biology approach.
AB - The possibilities of an in vitro translation system, based on cell components of the hyperthermophilic archaeon, Thermococcus kodakaraensis, to utilize at a lower temperature range were examined. Green fluorescence protein (GFP) was used as the reporter protein. Through optimization of several experimental conditions, the yield of active GFP production at 40°C increased 5-fold. The optimized system was encapsulated in liposome made by passing through the oil/water interfaces. The prepared liposomes were incubated at 40°C for 90 min, and fluorescence was examined by laser confocal microscopy. A significant increase in average fluorescence intensity was observed in liposomes with GFP mRNA, indicating that the T. kodakaraensis in vitro translation system is applicable for protein production within giant liposomes. These artificial cell models should provide the means to reconstitute various cell functions from a synthetic biology approach.
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U2 - 10.1109/MHS.2009.5351893
DO - 10.1109/MHS.2009.5351893
M3 - Conference contribution
AN - SCOPUS:77950941240
SN - 9781424450954
T3 - 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
SP - 176
EP - 181
BT - 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
T2 - 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
Y2 - 8 November 2009 through 11 November 2009
ER -