Open AccessT7 RNA polymerase‐driven inducible cell lysis for DNA transfer from Escherichia coli to Bacillus subtilis
Author(s) -
Juhas Mario,
Ajioka James W.
Publication year - 2017
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.12843
Subject(s) - t7 rna polymerase , bacillus subtilis , escherichia coli , lysis , biology , microbiology and biotechnology , recombineering , rna polymerase , dna , polymerase , bacteriophage , gene , biochemistry , genetics , bacteria
Summary 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. Reliable and efficient systems for the transfer of synthetic DNA between E. coli and B. subtilis are therefore of the highest importance. Using synthetic biology approaches, such as streamlined lambda Red recombineering and Gibson Isothermal Assembly, we integrated genetic circuits pT 7 L123, Repr‐ts‐1 and pL T 7pol encoding the lysis genes of bacteriophages MS 2, ΦX174 and lambda, the thermosensitive repressor and the T7 RNA polymerase into the E. coli chromosome. In this system, T7 RNA polymerase regulated by the thermosensitive repressor drives the expression of the phage lysis genes. We showed that 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 suitable for the efficient cell lysis and transfer of the DNA engineered in E. coli to other naturally competent hosts, such as B. subtilis .