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 proceedingConference 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 languageEnglish
Title of host publication20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
Pages176-181
Number of pages6
DOIs
Publication statusPublished - 2009 Dec 1
Externally publishedYes
Event20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science - Nagoya, Japan
Duration: 2009 Nov 82009 Nov 11

Publication series

Name20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science

Other

Other20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human Science
CountryJapan
CityNagoya
Period09/11/809/11/11

ASJC Scopus subject areas

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

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