Revision as of 23:15, 25 October 2007

Homemade E. coli S30

Cell-Free Systems (back to Intro)	Introduction
Chassis description	The Homemade E. coli S30 extract was prepared using E. coli strain BL21 (DE3) with loss of function mutations in OmpT endoproteinase and lon protease genes. This E. coli strain can express T7 RNA polymerase under the control of LacI, which is inducible by IPTG 1. The simple constitutive gene expression device BBa_T9002 has been used to characterize this cell-free chassis
Preparation protocol	Flow chart showing the protocol for making homemade E. coli S30 extract. The protocol is based on Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system by Kim DM et al 1.
Preliminary Testing

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 The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_T9002 BBa_T9002] has been used to characterize this cell-free chassis
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-|<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Homemade_E.coli_S30/Preparation_Protocol <font face=georgia color=#000066 size=4>Preparation protocol</font>]</center> ||
+|<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Homemade_E.coli_S30/Preparation_Protocol <font face=georgia color=#000066 size=4>Preparation protocol</font>]</center> ||[[Image:Homemade_E.coli_S30.png|thumb|400px|center]]Flow chart showing the protocol for making homemade ''E. coli'' S30 extract. The protocol is based on Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system by Kim DM et al <cite>1</cite>.
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 |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Homemade_E.coli_S30/Preliminary_Testing <font face=georgia color=#000066 size=4>Preliminary Testing</font>]</center> ||