Plasticity in chromosome number and testing of essential genes in Leishmania by targeting.

@article{Cruz1993PlasticityIC,
  title={Plasticity in chromosome number and testing of essential genes in Leishmania by targeting.},
  author={Angela Kaysel Cruz and Richard G. Titus and Stephen M Beverley},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={1993},
  volume={90 4},
  pages={
          1599-603
        }
}
We attempted to generate homozygous dhfr-ts (dihydrofolate reductase-thymidylate synthase) knockouts in virulent Leishmania major, an asexual diploid protozoan parasite. Transfection of a neo (neomycin phosphotransferase) targeting fragment yielded heterozygous replacement lines with high efficiency. However, second transfections with a hyg (hygromycin B phosphotransferase) targeting fragment in the presence of metabolites shown to rescue homozygous knockouts in attenuated Leishmania did not… CONTINUE READING
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