GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks

Kazuaki Yamaji; Tamotsu Kanai; Shin Ichiro M. Nomura; Kazunari Akiyoshi; Makiko Negishi; Haruyuki Atomi; Kenichi Yoshikawa; Tadayuki Imanaka

doi:10.1109/MHS.2009.5351893

GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks

Kazuaki Yamaji, Tamotsu Kanai, Shin Ichiro M. Nomura, Kazunari Akiyoshi, Makiko Negishi, Haruyuki Atomi, Kenichi Yoshikawa, Tadayuki Imanaka

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Abstract

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.

Original language	English
Title of host publication	20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
Pages	176-181
Number of pages	6
DOIs	https://doi.org/10.1109/MHS.2009.5351893
Publication status	Published - 2009 Dec 1
Externally published	Yes
Event	20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science - Nagoya, Japan Duration: 2009 Nov 8 → 2009 Nov 11

Publication series

Name	20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science

Other

Other	20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
Country	Japan
City	Nagoya
Period	09/11/8 → 09/11/11

ASJC Scopus subject areas

Computer Science(all)
Control and Systems Engineering
Electrical and Electronic Engineering

Access to Document

10.1109/MHS.2009.5351893

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Yamaji, K., Kanai, T., Nomura, S. I. M., Akiyoshi, K., Negishi, M., Atomi, H., Yoshikawa, K., & Imanaka, T. (2009). GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks. In 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science (pp. 176-181). [5351893] (20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science). https://doi.org/10.1109/MHS.2009.5351893

GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks. / Yamaji, Kazuaki; Kanai, Tamotsu; Nomura, Shin Ichiro M.; Akiyoshi, Kazunari; Negishi, Makiko; Atomi, Haruyuki; Yoshikawa, Kenichi; Imanaka, Tadayuki.

20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science. 2009. p. 176-181 5351893 (20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Yamaji, K, Kanai, T, Nomura, SIM, Akiyoshi, K, Negishi, M, Atomi, H, Yoshikawa, K & Imanaka, T 2009, GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks. in 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science., 5351893, 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science, pp. 176-181, 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science, Nagoya, Japan, 09/11/8. https://doi.org/10.1109/MHS.2009.5351893

Yamaji K, Kanai T, Nomura SIM, Akiyoshi K, Negishi M, Atomi H et al. GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks. In 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science. 2009. p. 176-181. 5351893. (20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science). https://doi.org/10.1109/MHS.2009.5351893

Yamaji, Kazuaki ; Kanai, Tamotsu ; Nomura, Shin Ichiro M. ; Akiyoshi, Kazunari ; Negishi, Makiko ; Atomi, Haruyuki ; Yoshikawa, Kenichi ; Imanaka, Tadayuki. / GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks. 20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science. 2009. pp. 176-181 (20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science).

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abstract = "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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AU - Negishi, Makiko

AU - Atomi, Haruyuki

AU - Yoshikawa, Kenichi

AU - Imanaka, Tadayuki

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