Abstract
The production and high level secretion of TaqI restriction endonuclease using bacterial secretion signal within the malE gene was achieved by cloning the PCR-amplified gene from Thermus aquaticus into E. coli. The maltose binding protein (MBP) part of the MBP-TaqI fusion protein expressed by this construct did not interfere with the biological activity of the TaqI restriction endonuclease. E. coli XL1 carrying pH185 produced 332 U ml−1 TaqI endonuclease 81% of which was secreted into the medium without apparent cell lysis. Optimization of culture conditions and selection of the host strain were found to be important for the efficient extracellular production of this protein.
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Toksoy, E., Özdinler, P., Önsan, Z. et al. High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli. Biotechnology Techniques 13, 803–808 (1999). https://doi.org/10.1023/A:1008938302312
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DOI: https://doi.org/10.1023/A:1008938302312