Tuning the Expression Level of a Gene Located on a Bacterial Chromosome

@article{Katashkina2005TuningTE,
  title={Tuning the Expression Level of a Gene Located on a Bacterial Chromosome},
  author={Joanna I. Katashkina and Alexandra Yu. Skorokhodova and Danila V. Zimenkov and Andrey Yu. Gulevich and Natalia I Minaeva and Vera G. Doroshenko and Irina V. Biryukova and Sergey V. Mashko},
  journal={Molecular Biology},
  year={2005},
  volume={39},
  pages={719-726}
}
A new method of constructing a set of bacterial cell clones varying in the strength of a promoter upstream of the gene of interest was developed with the use of Escherichia coli MG1655 and lacZ as a reporter. The gist of it lies in constructing a set of DNA fragments with tac-like promoters by means of PCR with the consensus promoter P tac and primers ensuring randomization of the four central nucleotides in the −35 region. DNA fragments containing the tac-like promoters and a selective marker… CONTINUE READING