cDNA cloning and heterologous expression of functional cysteine-rich antifungal protein Psd1 in the yeast Pichia pastoris.

@article{Almeida2001cDNACA,
  title={cDNA cloning and heterologous expression of functional cysteine-rich antifungal protein Psd1 in the yeast Pichia pastoris.},
  author={Marcius da Silva Almeida and Katia Maria Dos Santos Cabral and Luciano Neves de Medeiros and Ana Paula Valente and Francisca Cunha Almeida and Eleonora Kurtenbach},
  journal={Archives of biochemistry and biophysics},
  year={2001},
  volume={395 2},
  pages={199-207}
}
In the present work, we describe the cDNA cloning, expression in Pichia pastoris, purification, and characterization of the recombinant Pisum sativum defensin 1 (rPsd1), a novel Cys-rich protein presenting four disulfide bridges and high antifungal activity. Several parameters that affect the level of protein expression were assayed. The best condition yielded 13.8 mg/L (1.50 microg/10(8) cells) of active rPsd1. The recombinant rPsd1 was purified to homogeneity by cation exchange, followed by… CONTINUE READING