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T7 RNA polymerase-driven inducible cell lysis for DNA transfer from Escherichia coli to Bacillus subtilis.

last modified Oct 04, 2017 09:13 PM
Mario Juhas and Jim Ajioka from the Department of Pathology at the University of Cambridge have contributed to creating reliable and efficient systems for the transfer of synthetic DNA between E. coli and B. subtilis, supported by the SRI's SynBio Fund.

The majority of the good DNA editing techniques have been developed in Escherichia coli; however, Bacillus subtilis is better host for a plethora of synthetic biology and biotechnology applications. 

Using synthetic biology approaches, such as streamlined lambda Red recombineering and Gibson Isothermal Assembly, the team integrated genetic circuits  encoding the lysis genes of bacteriophages MS2, ΦX174 and lambda, the thermosensitive repressor and the T7 RNA polymerase into the E. coli chromosome.

In this system the T7 RNA polymerase regulated by the thermosensitive repressor drives the expression of the phage lysis genes. T7 RNA polymerase significantly increases efficiency of cell lysis and transfer of the plasmid and bacterial artificial chromosome-encoded DNA from the lysed E. coli into B. subtilis. The T7 RNA polymerase-driven inducible cell lysis system is therefore suitable for the efficient cell lysis and transfer of the DNA engineered in E. coli to other naturally competent hosts, such as B. subtilis.

The research obtained support from the SynBio Fund and OpenPlant Fund

The full article can be read here.

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