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From 2014.igem.org

Overview

In order to achieve the goal of integrating the directed evolution processes into a single automatic and controllable system, we come up with the following design.

1.The initiate state

M13 bacteriophage with gⅧ deficient carrying Prey sequence yet to be evolved. An E. coli with pBT-Bait and pRPT-gⅧ. At the temperature of 0 degree Celcius

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2.Infection

When the temperature rise to 30 degree Celcius, the M13 vector infects into the E. coli.

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3.Mutagenesis

When the M13 vector undergo replication, a library of different Prey is generated.

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4.Selection

Various Prey undergo selection processes with the help of Bacteria Two-Hybrid system

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5.Temperature regulated amplification

The temperature rise to 37 degree Celcius, so that mRNA of gⅧ can be translated into p8

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6. Rescue and Evolved

The p8 compensate M13 deficient, so that progeny phage can be generated

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